key: cord-0818905-28i8vkc9
authors: Hassanzadeh, Parichehr
title: The significance of bioengineered nanoplatforms against SARS-CoV-2: From detection to genome editing
date: 2021-03-04
journal: Life Sci
DOI: 10.1016/j.lfs.2021.119289
sha: 155a25823f1d115d7970427db50f0b6f3eca6769
doc_id: 818905
cord_uid: 28i8vkc9

COVID-19 outbreak can impose serious negative impacts on the infrastructures of societies including the healthcare systems. Despite the increasing research efforts, false positive or negative results that may be associated with serologic or even RT-PCR tests, inappropriate or variable immune response, and high rates of mutations in coronavirus may negatively affect virus detection process and effectiveness of the vaccines or drugs in development. Nanotechnology-based research attempts via developing state-of-the-art techniques such as nanomechatronics ones and advanced materials including the sensors for detecting the pathogen loads at very low concentrations or site-specific delivery of therapeutics, and real-time protections against the pandemic outbreaks by nanorobots can provide outstanding biomedical breakthroughs. Considering the unique characteristics of pathogens particularly the newly-emerged ones and avoiding the exaggerated optimism or simplistic views on the prophylactic and therapeutic approaches including the one-size-fits-all ones or presenting multiple medications that may be associated with synergistic toxicities rather than enhanced efficiencies might pave the way towards the development of more appropriate treatment strategies with reduced safety concerns. This paper highlights the significance of nanoplatforms against the viral disorders and their capabilities of genome editing that may facilitate taking more appropriate measures against SARS-CoV-2.

vaccines for targeting the hypervariable viruses. Synergistic application of the methods including the in silico and experimental approaches has been illustrated. Adapted from Ref. [27] .

Besides application for analyzing the empirical data and improving the efficiency of identification, in silico strategies can be used for predicting the immunogens for being included in new vaccine approaches based on epitopes [32] . Data obtained from the modular immune in vitro construct system and clinical studies have indicated that activation of all components of the immune system is required for inducing appropriate and long-lasting immune response [33] .

In general, vaccination efficiency depends on various factors such as the vaccine strain, nature of disease, proper schedule of vaccination, appropriate modeling approaches for predicting the immunization process performance, continuous monitoring particularly after presenting a novel vaccine, maintenance of high rates of immunization, or idiosyncratic reactions [34] . Regarding the development of effective and safe vaccines, various challenging issues should be addressed such as several stages of the life cycles of pathogens, diversity of pathogenic and physiologic oligomers and isoforms, host-related or vaccine attenuation failures, lack of appropriate response due to the various factors including the genetic ones, complexity of the immunopathology, epitope nature, identifying the pathological epitopes which could evoke specific antibody response, inappropriate or various degrees of immune responses or difficulties in prediction of this type of responses, and high rates of mutations in RNA viruses that may protect them against influenza [61] . Few clinical trials in China have represented favipiravir as an anti COVID-19 therapeutic agent due to the rapid clearance of virus and increased rate of improvement in patient's chest imaging [62] . This necessitates further confirmations by more vigorous and controlled trials.

Besides the viral target proteins, inhibitors which target the interaction of virus with host factors could be promising candidates (host-directed therapeutics) against the viral infections [63, 64] .

In general, drugs capable of direct targeting the virus appear to provide more therapeutic efficiency. SNG001, an experimental medication against the chronic obstructive pulmonary disorder is currently under evaluation in a phase-II clinical trial for its potential effectiveness against the COVID-19 [38] . Recently, enhanced morbidity and mortality in the hypertensive patients affected by COVID-19 has been reported [65] . Regarding the inhibitors of angiotensin converting enzyme (ACE) or antagonists of angiotensin receptor, there are no well-established studies demonstrating the potential harms or benefits of these drugs in COVID-19 patients [66, 67] . Despite the implication of ACE-2 in the viral entry (Fig. 2) and COVID-19 pathogenesis [68] , it could not be inhibited by ACE inhibitors which are usually prescribed in the clinic [69] . Journal Pre-proof ACE-2: angiotensin-converting enzyme 2. Adapted from Ref. [71] .

Furthermore, application of ACE inhibitors or antagonists of the angiotensin receptor may enhance ACE-2 expression and susceptibility of patients to the viral propagation and entry into the host cells [70] . These drugs have not influenced mortality or morbidity in 112 patients affected by both cardiovascular disease and COVID-19 [71] . Altogether, there is no appropriate scientific evidence or clinical trial data suggesting discontinuation of these medications in COVID-19 patients with co-existing cardiovascular disorders. Evaluating the accuracy of the contradictory hypotheses necessitates designing additional research projects.

EK1 and HR2-derived peptides as the inhibitors of entry/fusion appear to be effective against SARS-CoV-2 infections [72] . Furthermore, clustered regularly-interspaced short palindromic repeats (CRISPR)-Cas13d system has been suggested for targeting and cleaving the genome of SARS-CoV-2 [73] . Intracellular poly(ADP-ribose) polymerases can inhibit the replication of coronavirus and viral macrodomain has been suggested as an appropriate target for antivirus treatment [74] . Further evaluations are required for supporting these reports.

Based on the receptor-mediated endocytosis of coronavirus, targeting of endocytosis has been suggested as a promising approach against SARS-CoV-2 [75] . Over the last decade, huge amounts of costs and time for drug-related research and design has provoked development of more advanced strategies and techniques. Artificial intelligence (AI) technologies have offered Platforms powered by AI can be used for patient matching with relevant trials in the clinical settings leading to the improved cost-effectiveness and error reduction. A number of drugs targeting AP-2-associated protein kinase-1, as the host kinase capable of regulating clathrindependent endocytosis have been detected and baricitinib has been suggested as an appropriate drug against COVID-19 due to its inhibitory effect on the kinase activity [77, 78] . Inhibitors of p21-activated protein kinase-1, an enzyme which is involved in the viral entry and replication and can prevent micropinocytosis, have been suggested as therapeutic agents against COVID-19 [79, 80] . Noteworthy, inhibitors like caffeic acid, ketorolac, triptolide, or propolis are associated with low solubility and problems in cell penetrability [81] . In this respect, newer inhibitors such as minnelide and frondoside-A with improved solubility and potency have been developed [82, 83] that needs further evaluations.

The beneficial effects of corticosteroids for suppressing the hyper-inflammatory response in some COVID-19 patients has remained controversial. There is no well-established clinical evidence for supporting corticosteroid therapy against the lung injury associated with COVID-19 [84] . Recently, a retrospective study has shown the beneficial effects of low dose corticosteroids in the critically-ill COVID-19 patients [85] . Because of its high potency, dexamethasone may be considered for suppressing the hyperinflammatory or immunologic reactions in patients with severe conditions, however, after appropriate infection control. Furthermore, dexamethasone

Preventing activation of Fc receptors may also be helpful for reducing the inflammatory response induced by SARS-CoV-2 [86] . Furthermore, blocking interleukin-6 (IL-6) receptors or granulocyte-macrophage colony stimulating factor may reduce SARS-CoV-2-induced immunopathology [87] .

In the case of an infection for which there is no particular treatment approach, CP therapy (providing specific antibodies with human origin) has been suggested as a promising strategy [88] . Regarding COVID-19, CP can be obtained from a recovered patient with humoral immunity against the disease [89] . However, evaluation of the safety and efficiency of CP therapy has remained challenging that may be due to imprecise mechanism of action or inappropriate clinical trials [88] .

mAbs can be applied for direct attacking and neutralizing the virus, preventing the infection of host cells, or blocking the spike proteins for preventing virus attachment to the host cells [90] .

In patients recovered from the infections of COVID-19, reproducing or engineering of the functional copies may provide antibodies for mimicking or increasing the immune system attack against the SARS-CoV-2 [91] . Tocilizumab as a mAb against the receptor of IL-6 which has received FDA approval for treating patients with giant cell arteritis and idiopathic or rheumatoid arthritis [92] , has been recently applied as the immunosuppressive drug in COVID-19 patients in critical conditions in Italy and China and promising results have been reported [93] . Meanwhile, performing various clinical trials in different countries is required for obtaining more conclusive data.

J o u r n a l P r e -p r o o f Journal Pre-proof TE techniques which can be applied for repairing or replacing the damaged tissues and organs, improving the efficiency of traditional treatment strategies against a variety of disorders, screening of drugs, or immunomodulation [94, 95] , may also be useful against the viral outbreaks via designing the viral models, vaccine platforms, and systems for delivery of therapeutics [96] .

Tissue-engineered lung models can be used for evaluating the developmental process of tissue and its physiological or pathological conditions including the infections induced by viruses [97] .

However, TE may be associated with a variety of limitations that necessitates application of the modeling approaches and newer technologies including 3D printing for obtaining biocompatible supporting materials [98] .

Drug repurposing with present antivirals is also an encouraging approach for treatment of the viral infections [99] . Noteworthy, the safety and efficiency of the aforementioned investigative, preventive, and treatment strategies including the traditional medicines or their combination with conventional drugs against SARS-CoV-2 have not been fully approved by health authorities [100, 101] . Indeed, there is no therapeutic agent with absolute safety. Development of the efficient preventative or treatment strategies for limiting infections necessitates consideration of the unique properties of pathogens including the viruses, selection of the appropriate sample size, sharing of datasets extracted from high-quality evaluations, and identifying the limitations of studies that might result in the increased efficiency and reduced safety concerns.

activator-like effector, zinc finger, or mega-nucleases), and clustered regularly-interspaced short palindromic repeats system have been selected as the breakthrough and methods of the year by the prestigious journals, Nature methods and Science [103, 104] . These nucleases induce double strand breaks (DSB) in the genes in a site-specific manner. Creation of DSB at specific sites is of critical importance in editing of genes (Fig. 3) . The breaks can be repaired via the homologous or non-homologous pathways leading to the targeted-mutations [102] . In general, editing of the genome depends on the mechanics of DSB repair through which various enzymes join DNA ends or a template (homologous sequence) is applied to regenerate the missing sequence of DNA at break point. In this context, a vector is created with appropriate eradication and enable taking genes from the human cells or replacing genes capable of activating cancer or other undesired defects. Because of its efficiency and precision, CRISPR can be applied for establishment of the large-scale productions [108] .

During decades, mice have been used as a disease model host. Using CRISPR system enables bridging the gaps between the animal disease models and clinical trials via creation of the transgenic models in large animals and non-human primates [109, 110] , targeting and eliminating the endogenous retroviruses leading to the reduced risk of disease transmission and immune barriers [108] . CRISPR-Cas9 as the most versatile technique for engineering or editing of the genomes and gene therapy, has enabled direct introducing of sgRNA and Cas9 protein into the zygotes for obtaining intended modifications of genes, bypassing cell targeting stage in the generation of transgenic lines, and significant reduction of generation time [111] . The most widely applied CRISPR-Cas9 machinery has been derived from the Streptococcus pyogenes and Cas9 endonuclease from F. novicida is capable of RNA targeting [112] . CRISPR-Cas9 system of F. novicida has been applied for targeting single-stranded RNA (+ssRNA) virus genome (hepatitis C virus, HCV) [113] . sgRNA and Cas9 targeted to 5' and 3' un-translated regions of HCV genome led to the significant reduction (50-60%) of the expression of viral protein [113] .

These findings might be helpful for development of resistance to RNA viruses using the technology of gene editing. genome and showed capability of simultaneous engineering of 2 viral genes that might significantly expand biomedical applications of the Vaccinia virus as a promising vector for a variety of vaccines against the infectious diseases, delivery of genes, and anticancer immunotherapies [117, 118] . In usual, CRISPR-Cas system targets DNA in hosts or viruses but the system is also capable of RNA targeting and editing [114, 119] . RNA-targeting CRISPR-Cas effector C 2 C 2 system via cleavage of the viral RNA and preventing the replication of virus can be considered as a promising strategy against RNA viruses [120] .

Acquiring a better understanding about the host response to SARS-CoV-2 might be of therapeutic significance. DNA or RNA editing by the endogenous deaminases can restrict the replication of particular viruses [121] . Following SARS-CoV-2 virus entry into the human body, two deaminase enzymes have shown the capability of editing the RNA of virus and affecting its replication [121, 122] that might be of great significance against COVID-19. APOBECs and ADARs are deaminase families which are expressed in mammals and change nucleotide building blocks of RNA in the virus via amino group removal [123] . They are implicated in gene editing of the coronavirus that may influence the fate of virus and affected patient [122] . After singlenucleotide variant analysis in RNA sequencing data sets from the extracted fluid from the COVID-19 patient's lung, low levels of the mutation occurence were observed and nucleotide alterations were identified that may be due to the editing of RNA [122] . By comparing the defense [124] . Following application of CRISPR-Cas machinery for producing DNA virusresistant plants [125] , CRISPR-Cas9 system has been reprogrammed for conferring immunity against RNA viruses in Arabidopsis and Nicotiana benthamiana plants leading to the reduced accumulation of the viral RNA and symptoms of infection [126] . Transgenic plants demonstrated inheritable resistance and a remarkable reduction of viral accumulation was observed in the progenies indicating the importance of CRISPR-Cas9 technology for producing plants with stable resistance to RNA viruses [127] . Such findings could be applicable to other species for direct targeting RNA viruses in the eukaryotes and preventing further infections.

Nucleases which can be programmable enable accelerated engineering of the genomes in cells or organisms [128], however, their targeted delivery may be quite challenging. In this respect, Cas9-sgRNA ribonucleoproteins-loaded murine leukemia VLPs have been applied for inducing efficient editing of genomes in a variety of primary cells and cell lines [129] .

Application of targeted-RNA recombination has been shown as a promising approach for manipulating the genome of coronavirus (specially in 3´ part) [130] . [112] . This can be done via introducing RNA-guided nucleases into the host cells during the viral replication leading to the inhibition of replication [132] . Engineered replication defective viruses may also be applied for gene editing. In this sense, viral vectors as delivery devices for nucleases and DNA templates have been considered as useful tools in the settings of genome editing [133] . Lentiviral vectors can permanently induce genetic modifications in the target cells because of the integrase dependent mechanisms [134] . This type of mechanisms are of critical significance for inducing stable genetic deficiencies in target cells [135] . Using adenoviral vectors for inducing Apc mutations or chromosomal inversion or rearrangement represent these vectors as the donor DNA template source for genome editing in a homologydirected manner and promising platforms which are able to introduce designer nucleases in vivo [136] . However, activation of the adaptive and innate immune responses against the viral particles or presence of the designer nucleases for long periods of time in target tissues are challenging issues [137] which should be addressed. Ideally, editing of genomes should be performed in accurate and rapid manner for limiting the potential toxicities and off target effects because of the expression of effectors in a sustained fashion [138] . In order to minimize the limitations of the genome editing, enhancement of the specificity and safety of the engineered nucleases and improved detection capability of off target effects, obtaining a deeper knowledge Imminent breakthroughs in nanotechnology have revolutionized theranostic strategies.

Functional system engineering at molecular scales, designing nanovectors including the nanoparticles (NPs) capable of loading a variety of imaging or therapeutic agents for early detecting the transforming cells or diseases, targeted therapy, and treatment outcome monitoring [141, 142] Hybrid nano-biosystems containing virus-derived biomolecules conjugated to the NP surface which can serve as specific probes for virus detection are more advantageous over the conventional methods in terms of specificity, stability, sensitivity, and performance [144] .

Journal Pre-proof Nanocarriers due to their suitable physicochemical characteristics such as small size of particles, large surface area, and tunable charge of the particle surface which facilitate particle penetration across the cell membrane, improved solubility, stability, and bioavailability of the entrapped drugs, targeted delivery, and reducing drug resistance can be suitable reservoirs for antiviral agents [145] . A variety of liposomal formulations, polymeric NPs, nanosuspensions, nanoemulsions, dendrimers, solid lipid NPs, and niosomes have been presented against the viral infections [146]. Furthermore, graphene oxide and silver nanocomposites has shown promising antiviral activities [147] . Silver NPs (Ag-NPs) by interacting with cell surface receptors inhibit virus entry into the host cells and prevent its reproduction [148] . They interact with the genome of double-stranded RNA viruses and inhibit their replication [149] . Ag-NPs undergo color alterations because of the localized surface plasmon resonance effect [150] . Ag-NPs modified by mercaptoethane sulfonate or tannic acid have been shown to prevent infections induced by type 1 or 2 of the herpes simplex virus via inhibiting the penetration, attachment, and spread of virus [151, 152] . Gold NPs (AuNPs) because of their distinctive electric, catalytic, and photonic characteristics and capability of interactions with a variety of biomolecules such as RNA aptamers, single-stranded DNA, and antibodies can be applied in the viral detection settings [153] . For virus detecting, Au-NPs are capable of performing various functions such as amplification of color, fluorescence enhancing or quenching, and light scattering. Bioconjugates of Au-NPs can be applied in the electrochemical, colorimetric, fluorometric, and scanometric systems for detecting human viral groups [154] , (Fig. 6) .

detection. These nanoparticles with high sensitivity and specificity are capable of transducing various signal types including the electrical, optical, and catalytic signals. They could also be applied as the scaffolds for bio-immobilization. Adapted from Ref. [153] .

Highly sensitive and simple Au-NPs-based assays have represented Au-NPs as promising nanoplatforms for detecting of various virus types [155] . Meanwhile, special attention should be paid to the application of advanced and rigorous methods for synthesis of the stable and uniform Au-NPs with several surface modification possibilities, appropriate conjugation of the specific targeting biomolecule to the NP surface, application of the negative and positive controls and reference tests, and in-field assessment of the novel Au-NPs-based techniques for confirming their specificity and sensitivity. In recent years, a highly selective colorimetric analysis has been performed for identifying the viral lysine using the molecular-driven Au nanorods [156] .

Carbon nanotubes (CNTs) with enormous potentials in a variety of scientific domains such as nanomedicine, can be applied as nanoreserviors for targeted delivery and controlled release of a J o u r n a l P r e -p r o o f variety of therapeutics. CNTs with superior biocompatibility, solubility, mechanical characteristics, and thermo-electrical conductivity have attracted a growing interest in theranostic fields such as high-resolution imaging and bio-sensing [157] [158] [159] [160] [161] [162] . Besides the viral detection, CNTs can be applied as nanocarriers of antiviral agents and improve their efficiency. For instance, chemical linkage of ribavirin or isoprinosine on the surface of single-walled CNTs for carrying the drug across the cell membranes has led to the improved drug efficiency [163, 164] .

Besides Au-NPs and mesoporous silica NPs, CNTs can be used as CRISPR-Cas9 carriers for delivery and editing of genomes. CNTs-DNA capable of coding Cas9-protein and guide RNA is a suitable nanosystem for editing specific genomes [165] . Quantum dots capable of emitting the photons with specific wavelengths, can provide highly-sensitive and robust fluorescence perspective for point-of-care viral detection [166, 167] . These semiconductor particles may be modified and along with various nanostructures can be applied for virus detecting [166] .

As aforementioned, safe and efficient delivery of the elements of gene editing towards the specific region and providing targeted correction of genes devoid of off-target adverse effects may be quite challenging. In this context, various strategies have been proposed for improving the delivery efficiency and facilitating clinical translation of the technology of genome editing [168] . Engineering the non-viral NPs including the liposomes and polymeric, inorganic, or lipid NPs enables overcoming many of the limitations associated with the physical or viral approaches for delivery of CRISPR-Cas9 and attenuating off-target effects [168] , (Fig. 7) .

Journal Pre-proof Based on the mRNA activity in the cytosol, it can prevent the risk of mutagenesis in the genome [174] . Despite the potentials of mRNA for preventing or treating a wide range of disorders and development of advanced in vitro transcription technology for controlling the immunogenicity and stability of mRNA, extensive applications of mRNA-based drugs necessitates designing the NPs and stability of NPs was evaluated using various concentrations of the cryoprotectants; mannitol, trehalose, or sucrose. It has been found that addition of trehalose or sucrose (5%, w/v) to the lipid NPs provides a suitable efficiency of mRNA delivery for about 3 months [181] .

Microfluidic mixing process is a common technique for mRNA encapsulation in lipid-NP systems [182] . Microfluidic mixing method can be applied for development of appropriate lipid-NP systems capable of encapsulating the negatively-charged macromolecules such as gold NPs, mRNA, and plasmids. Lipid-NPs composed of the cationic lipid, PEG, cholesterol, distearoylphosphatidylcholine (DSPC), and siRNA could be appropriately constructed using the techniques of microfluidic mixing.

Molecular simulations have revealed the nano-structured core of the lipid-NPs with aqueous compartments holding siRNA. Besides the imaging applications, lipid-NP-siRNA systems could be used as promising therapeutics [182] . In recent years, microfluidic techniques have been applied for improving the reproducibility and quality of lipid NPs leading to more efficient tissue delivery of siRNA [183, 184] . Formation of lipid-NPs based on the fusion-dependent procedure provides a deeper knowledge about the formation mechanism and structures of the lipid-NPsnucleic acid complex that might facilitate development of more potent lipid-NP-nucleic acid polymers for biomedical applications including the gene therapy [185] .

As aforementioned, traditional therapeutics and vaccines against the viral disorders may be associated with a variety of side effects including the toxicity, resistance, and high costs. Over the last decade, increasing interest has been attracted towards the mechanism and application of RNAi as a tool for manipulation or controlling the expression of genes and inhibiting viral replication [186, 187] . RNAi is recognized as one the most efficient and specific methods for silencing the genes including the target transcripts of mRNA [188] . RNAi induced by the small-J o u r n a l P r e -p r o o f Journal Pre-proof interfering RNA (siRNA) is capable of inhibiting the viral antigen expression and providing novel antiviral treatment approaches [189] . Because of the capability of gene silencing, activity in the cellular processes, and low cost, siRNA has been considered as an attractive medicinal modality. Simultaneous treatment with various shRNAs or siRNAs has been suggested against the viral infections and mutations [190] .

Noteworthy, natural RNA may be associated with various challenging issues such as nonefficient delivery to the target tissues, short half-life because of the susceptibility to the nucleases, rapid clearance, immune response stimulation, and off target effects. Furthermore, specific targets are required for combating against the infections induced by viruses and avoiding the mutant escapes [191] . In general, clinical translation necessitates enhanced stability and reduced toxicity of RNAs, targeted delivery, and preventing mutant escapes. Modification of RNAs may result in the reduced immunogenicity and viral escape and enhanced stability, however, the modified RNAs may still demonstrate poor uptake into the cells because of their negative charge, hydrophilicity, and rapid clearance [192] . Failure of the synthetic siRNAs in crossing the biomembranes via the passive diffusion that may be due to their polyanionic nature and high molecular weights, has provoked the need to develop the technologies for transmembrane delivery of drugs [193] . In this respect, various delivery systems including the lipid NPs as one of the most advanced nanoplatforms for delivery of therapeutic agents including the biomolecules, have been designed for providing efficient uptake into the cells and accumulation at the target site or synergistic effects between the traditional drugs and RNAs for preventing the resistance [194] . Besides the ease of manufacturing and safety, lipid NPs can be functionalized for achieving proper tropism that might be of great significance when RNA interference is not organ-specific [195] . In recent years, the technology of lipid NPs has J o u r n a l P r e -p r o o f Journal Pre-proof facilitated clinical translation of nanodrugs such as those for efficient delivery of siRNA against a variety of disorders such as amyloid polyneuropathy that prevented production of pathological proteins. Development of Onpattro, a lipid NP-based siRNA drug, has paved the way for clinical translation of the lipid NP nanomedicines which contain nucleic acids for expressing or silencing the target genes [196, 197] . Increasing research efforts have been attracted towards the maximizing siRNA-lipid NPs potency for gene silencing [198] . Cationic lipid NPs are promising nanosystems for delivery of siRNA against the RNA virus, HCV. Intravenous treatment has reduced core protein of the virus in transgenic mice [199] . Increasing clinical trials have also been carried out for evaluating siRNA-loaded NPs including the lipid-based ones against the infections induced by a variety of viruses such as HBV, HIV, and HCV [200] . siRNAs entrapped into the lipid NPs have successfully targeted Ebola virus outbreak strain and provided remarkable protection and fully recovery for the rhesus monkeys when the therapy was begun three days after the exposure [201] . In this sense, lipid NPs-delivered siRNAs have been suggested for counteracting this type of lethal viral disease in humans.

RNA vaccines can also be packaged within the lipid NPs as vectors [202] . This type of vaccines are currently under development for fighting against the coronavirus pandemic. mRNA vaccines could be suitable substitutes to the traditional vaccine technology that may be due to their high potency and short cycles of production [203] . mRNA vaccine entrapped in the lipid NPs can generate vigorous immune responses [204] . It is possible to improve the tolerability of mRNA vaccine without influencing its potency [205] .

Combination of the lipid NP-based delivery system and sgRNA has provided a significantly increased activity in vivo in both rat and mouse models. It has been shown that lipid NP-based system is capable of delivery the components of CRISPR-Cas9 for obtaining the clinically-J o u r n a l P r e -p r o o f Journal Pre-proof relevant genome editing levels in vivo and significant reduction of transthyretin. Lipid NP-based delivery by achieving > 97% knockdown of the target protein after single dose and providing cumulative level of editing after multiple doses has represented lipid NP system as an effective platform for editing the genomes. The biodegradable and transient lipid NPs-based system containing poly(amidoamine), chitosan, or cell-penetrating peptides for CRISPR-Cas9 delivery has been well-tolerated and the levels of editing was preserved for about one year [206, 207] indicating the ability of lipid NPs for providing knockdown and genome editing with high durability. Noteworthy, durability is an important factor which determines the efficiency of any gene editing process [208] . In general, engineering of lipid NPs for regulation of genes in specific cells or tissues is of great importance as the technology could provide the possibility of gene therapy in an organ-specific fashion [205] . Using the selective organ targeting (SORT) approach which is compatible with various techniques of genome editing and may be helpful for designing protein-replacement or gene correcting therapeutic agents in targeting organs, lipid NPs carrying the therapeutic agents based on the nucleic acids have been recently bioengineered for inducing lung-, liver-, or spleen-specific regulation of genes [209] . Cell-or organ-targeted SORT lipid NPs could be engineered for selective editing of the hepatocytes, endothelial cells, epithelial cells, T cells, or B cells [209] . Administration of tissue-specific mRNA-encapsulated lipid NPs has provided sustained production of the therapeutic proteins with minimal safety concerns [205] . In a study conducted by Cheng et al., i.v. treatment with SORT lipid-NPs led to Synthetic NP vectors composing of the nucleic acids poly(bamino esters) have been shown promising for delivery of nucleic acids against a variety of diseases [210] . For expanding this advantage for systemic mRNA delivery, hybrid lipid-polymer NPs have been developed to deliver mRNA into the lungs [211] . Co-formulation of poly(β-amino esters) with polyethylene glycol-lipid has led to the development of mRNA formulations with enhanced stability and potency and capable of mRNA delivery to the lungs in mice following intravenous injection In general, the molar ratios and components of lipid NPs which usually consist of the cholesterol, phospholipids, polyethylene glycol lipids, and cationic lipids, have been optimized for ensuring efficient delivery of the nucleic acids to the tissue and providing potent silencing of genes at clinically-relevant doses [212] . Increasing the molar components of lipid NPs with additional molecules for tuning NP internal charge can facilitate the delivery of therapeutics in an organspecific fashion and affect the cellular fate of NPs [213] . Addition of the cationic lipid has been shown necessary for increased efficiency of SORT-lipid NPs [209] . Using SORT-lipid NPs has also enabled CRISPR-Cas system-based genome editing in a tissue-specific fashion [209] .

Generating SORT-lipid NPs holding Cas9 mRNA and guide RNA facilitated editing of genes in the extra-hepatic tissues. Using various formulations of SORT-lipid NPs in mice has enabled selective inducing of target genome editing in a tissue-specific manner [209] . However, the precise mechanisms by which the lipid NPs can be directed towards the specific organs by SORT moderation have remained to be addressed. Identifying the underlying mechanisms would enable fine tuning of tissue-or organ-specificity of lipid NPs. Furthermore, the probable immunogenicity should be rigorously evaluated prior to the implementation of SORT technique J o u r n a l P r e -p r o o f Journal Pre-proof for rational designing of lipid NPs for therapeutic modification of genes in various organs [207, 209] .

Using graphene oxide-or arginine-modified nanostructures or those consisting of Cas9-peptide for CRISPR/Cas9 delivery has provided suitable editing efficiency. NPs containing graphene oxide-PEG-polyethyleneimine are capable of protecting the cargo, sgRNA-Cas9, against the enzymatic hydrolysis [213, 214] . Development of a customizable nanocapsule for delivery of the single-guide RNA (sgRNA) and Cas9 has enabled controlled stoichiometry of the components of CRISPR and efficient generation of targeted genome edits, and limited the potential safety concerns. This nanoplatform has efficiently delivered CRISPR-ribonucleoprotein complex for somatic genome editing in vitro and in vivo [214, 215] .

As aforementioned, coronaviruses including SARS-CoV and MERS-CoV are pathogens capable of causing widespread pneumonia outbreaks with high mortality and morbidity rates. Highly contagious nature and clinical importance of these viruses necessitate application of advanced detection techniques. Using Au-NPs for detecting coronaviruses is based on the development of specific and rapid detection of molecules via colorimetric and electrochemical assays [153] .

These assays particularly the colorimetric one put an end to the requirement of skilled personnel or complex instrumentation and provides negative or positive results only in the liquid phase which can be easily identified (within 5 min) by unveild eye. Using the technique enables detecting target SARS virus nucleic acids with the sensitivity limit of about 100 fM [153] indicating the appropriateness of technique for early virus diagnosis that might be of great significance for these highly contagious viruses. siRNA ability for prophylaxis or therapy of the coronavirus-related infections has been previously evaluated [216] . siRNA pre-existed in the host cells is capable of inhibiting the J o u r n a l P r e -p r o o f Journal Pre-proof replication of SARS-CoV and further infections because of disrupting virus RNA and inactivating the replication machinery of the virus [217] . siRNA duplexes have been assessed for anti-SARS-CoV effects in the primate cells and active duplexes and showed prolonged inhibitory effects on the replication of SARS-CoV and further infection. Combination of the active sequences provided increased antiviral potency and reduced viral escape because of the mutation in siRNA target. In this sense, integrating siRNA duplexes has been shown as a promising approach for development of the antiviral therapeutics [217] . Application of siRNA duplexes for treating patients infected by CoV necessitates efficient delivery of this type siRNAs to the lungs.

In the Rhesus macaque affected by SARS coronavirus, siRNA has reduced viral load and replication and protected lungs against the diffuse alveolar damage. It has been represented as a safe biological agent with enormous potentials in targeted therapy or prophylactic antiviral regimens [218] . Liver is one of the most vulnerable organs to the virus attacks. Patients affected by the liver diseases may be more vulnerable to the negative outcomes of COVID-19 [221] . Based on the reports, dendrimer-based lipid NPs have effectively delivered miRNAs/siRNAs for normalizing the functions of liver [222] . mRNA-encapsulated lipid NPs can be applied for production of the therapeutic proteins and gene-editing complexes for correcting disease-induced mutations in the hepatocytes [223] . Single treatment with a lipid NP-based delivery system has led to an efficient editing of transthyretin gene in mouse liver and a significant and durable reduction of protein J o u r n a l P r e -p r o o f Journal Pre-proof contents in the serum [223] . Nanoblades which can be used for homology-directed repairs, are capable of ribonucleoprotein cargo delivery in a rapid and transient fashion, mediating knock-in in various cell lines and genome-editing in a dose-dependent manner without significant influence on target cell proteome, programming with Cas9 proteins for mediating transient targeted gene activation or up-regulation [129] . Viral-derived Cas9-sgRNA ribonucleoproteinsloaded nanoblades have been capable of editing the genomes in mouse liver [129] .

Application of AI techniques for accelerating the process of nanofabrication and solving the associated problems, developing smarter and hybrid technologies, producing the nanoarchitectures with enhanced power of computation, and evaluating the impact of nanostructures on the biosystems [224] enable overcoming the potential limitations of the nanotech-based approaches. Computational modeling is an essential step for creating functional systems for delivery of nanotherapeutics and their clinical translation. Acquiring an improved understanding about the underlying mechanisms of the bio-events and bio-nano interactions and predicting the effects of formulation factors on distribution or delivery of the entrapped agent and dose-response are of critical importance for designing the nanotherapeutics with increased efficiency of targeting and reduced safety concerns. Patient-specific models capable of providing special opportunities can be applied for personalized theranostics [225] [226] [227] . In recent years, advancements in the structural and molecular biology and computational genomics have provided a deeper knowledge about the structures of protein targets in both virus and host that might result in the development of more efficient antiviral agents [228] . Computer simulations via application of the structural information and mutational analyses facilitate rational design of new antiviral agents including the nucleotide inhibitors [229] . Advancements in translational J o u r n a l P r e -p r o o f Journal Pre-proof bioinformatics, structural biology, pharmacogenomics, in silico techniques, and virtual designing of ligands enable overcoming the newly-emerged and highly-mutating viruses and development of vaccines and a variety of highly-specific antiviral drugs including those capable of inhibiting the proteins of virus [230, 231] . Computationally-designed gRNAs targeting Vaccinia virus genes have facilitated evaluating the functions of genes [118] . Inverse computational fluid dynamics (CFD) modeling enables identification of the contaminants and their spread [232] . CFD models can be applied for investigating the coronavirus spread [233] . Using computational technique, a project has been recently designed (Fig. 10) to construct COVID-19 disease map for providing a comprehensive knowledge about the mechanisms of interactions between SARS-CoV-2 virus and host and development of models with high-quality capable of linking to the data repositories [234] . host, recovery of the host cells, the mechanisms of repair, and immune responses [234] .

Obtaining the improved knowledge of disease which is associated with complex pathomechanisms and multi-organ and multi-cellular nature of the infection, and facilitating the design process of efficient theranostics are other beneficial aspects of the presented map [234] .

Various pathways are included in the map such as the replication cycle of virus and mechanisms of transcription, the effect of SARS-CoV-2 on the pulmonary blood pressure, heme catabolism, apoptosis, interferon 2 signaling, and SARS-CoV-2 proteins. COVID-19 diagram collections and metabolic model of the alveolar macrophages in a patient affected by the virus are also incorporated in the map [234, 235] . Combination of the illustrative representations of the mechanisms of COVID-19 with underlying models in the map provides a suitable plan for the virologists, immunologists, and clinicians for collaboration with the computational biologists and data analyzers in order to build rigorous models and accurate interpretation of data [236, 237] . It would also be possible to better understand host susceptibility characteristics including the age and gender, progression of disease, mechanisms of defense, and treatment response. Application of the map with other disease maps enables assessment of the comorbidities [234] .

As aforementioned, the ability of precise genome modification and regulating particular genes might be of great significance in biomedicine. In order to better characterize the variables which control off-target or on-target activity of the Type II CRISPR-Cas9 system, a biophysical model

Structural definition of a neutralization-sensitive epitope on the MERS-CoV S1-NTD

SARS-CoV-2 and SARS-CoV spike-RBD structure and receptor binding comparison and potential implications on neutralizing antibody and vaccine development

Structure of SARS coronavirus spike receptor-binding domain complexed with receptor

Proteolytic activation of the SARS-coronavirus spike protein: cutting enzymes at the cutting edge of antiviral research

Structural definition of a unique neutralization epitope on the receptor-binding domain of MERS-CoV spike glycoprotein

History and recent advances in coronavirus discovery

Virus isolations from common colds occurring in a residential school

A Novel Coronavirus from Patients with Pneumonia in China

Hosts and Sources of Endemic Human Coronaviruses

The continuing 2019-nCoV epidemic threat of novel coronaviruses to global health-The latest 2019 novel coronavirus outbreak in Wuhan

Potential impact of seasonal forcing on a SARS-CoV-2 pandemic

The trinity of COVID-19: immunity, inflammation and intervention

Coronavirus (COVID-19) update: Serological tests. Food and Drug Administration

Assay Techniques and Test Development for COVID-19 Diagnosis

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

A Multipeptide vaccine is safe and elicits T-cell responses in participants with advanced stage ovarian cancer

Protein Engineering Strategies for the Development of Viral Vaccines and Immunotherapeutics

Peptide-based synthetic vaccines

Design of an epitopebased peptide vaccine against spike protein of human coronavirus: an in silico approach. Drug Design

Peptide-based vaccines: Current progress and future challenges

Peptide-Based Vaccinology: Experimental and Computational Approaches to Target Hypervariable Viruses through the Fine Characterization of Protective Epitopes Recognized by Monoclonal Antibodies and the Identification of

Epitope-based vaccine against influenza

A cross-protective mAb recognizes a novel epitope within the flavivirus NS1 protein

A J o u r n a l P r e -p r o o f Journal Pre-proof potential role for monoclonal antibodies in prophylactic and therapeutic treatment of influenza

Epitope mapping: the first step in developing epitope-based vaccines

In silico analysis of six known leishmania major antigens and in vitro evaluation of specific epitopes eliciting HLA-A2 restricted CD8 T cell response

How to design effective vaccines: lessons from an old success story

Vaccine programmes and policies

Primary vaccine failure to routine vaccines: Why and what to do?

Engineered Dengue Virus Domain III Proteins Elicit Cross-Neutralizing Antibody Responses in Mice

Progress and prospects on vaccine development against SARS-CoV-2. Vaccines

Comparison of DNA and mRNA vaccines against cancer

Advances in mRNA Vaccines for Infectious Diseases

Adenoviral vector platform for transduction of constitutive and regulated tricistronic or tripletranscript transgene expression in mammalian cells and microtissues

Characterization of chikungunya virus-like particles

Virus-like particles in picornavirus vaccine development

Longitudinally profiling neutralizing antibody response to SARS coronavirus with pseudotypes

Mechanism of immunopotentiation and safety of aluminum adjuvants

Fluad(R)-MF59(R)-Adjuvanted Influenza Vaccine in Older Adults

Adjuvant system AS01: Helping to overcome the challenges of modern vaccines

Comparative Safety of Vaccine Adjuvants: A Summary of Current Evidence and Future Needs

Don't rush to deploy COVID-19 vaccines and drugs without sufficient safety guarantees

The spike protein of SARS-CoV-a target for vaccine and therapeutic development

Anti-spike IgG causes severe acute lung injury by skewing macrophage responses during acute SARS-CoV infection

Chloroquine and hydroxychloroquine in the treatment of COVID-19 with or without diabetes: A systematic search and a narrative review with a special reference to India and other developing countries

SARS-CoV-2 and the Use of Chloroquine as an Antiviral Treatment

Pharmacology of Chloroquine and Hydroxychloroquine

Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial

Compassionate use of remdesivir for patients with severe COVID-19

Remdesivir is a direct-acting antiviral that inhibits RNAdependent RNA polymerase from severe acute respiratory syndrome coronavirus 2 with high potency

Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial

A trial of lopinavir-ritonavir in adults hospitalized with severe COVID-19

Synergistic combinations of favipiravir and oseltamivir against wild-type pandemic and oseltamivir-resistant influenza A virus infections in mice

Experimental Treatment with Favipiravir for COVID-19: An Open-Label Control Study

Rational Design of Novel Highly Potent and

Overlapping and discrete aspects of the pathology and pathogenesis of the emerging human pathogenic coronaviruses SARSCoV, MERS-CoV, and 2019-nCoV

SARS-CoV2: should inhibitors of the renin-angiotensin system be withdrawn in patients with COVID-19?

Association of Inpatient Use of Angiotensin Converting Enzyme Inhibitors and Angiotensin II Receptor Blockers with Mortality Among Patients With Hypertension Hospitalized With COVID-19

A pneumonia outbreak associated with a new coronavirus of probable bat origin

Evaluation of angiotensinconverting enzyme (ACE), its homologue ACE2 and neprilysin in angiotensin peptide metabolism

COVID-19 and Angiotensin-Converting Enzyme Inhibitors and Angiotensin Receptor Blockers: What Is the Evidence?

Outcomes in patients with

Virus against virus: A potential treatment for 2019-nCov (SARS-CoV-2) and other RNA viruses

The coronavirus macrodomain is required to prevent PARP-mediated inhibition of virus replication and enhancement of IFN expression

SARS coronavirus entry into host cells through a novel clathrin-and caveolae-independent endocytic pathway

The significance of artificial intelligence in drug delivery system design

AP-2-associated protein kinase 1 and cyclin G-associated kinase regulate hepatitis C virus entry and are potential drug targets

Baricitinib as potential treatment for 2019-nCoV acute respiratory disease

Role of the p21-activated kinases (PAKs) in influenza A virus replication

3-Triazolyl ester of Ketorolac: A "Click Chemistry"-based highly potent PAK1-blocking cancer-killer

Phosphonooxymethyl Prodrug of Triptolide: Synthesis, Physicochemical Characterization, and Efficacy in Human Colon Adenocarcinoma and Ovarian Cancer Xenografts

Frondoside A from sea cucumber and nymphaeols from Okinawa propolis: Natural anti-cancer agents that selectively inhibit PAK1 in vitro

Clinical evidence does not support corticosteroid treatment for 2019-nCoV lung injury

Potential benefits of precise corticosteroids therapy for severe 2019-nCoV pneumonia

Understanding SARS-CoV-2-mediated inflammatory responses: From mechanisms to potential therapeutic tools

Aberrant pathogenic GM-CSF+ T cells and inflammatory CD14+CD16+ monocytes in severe pulmonary syndrome patients of a new coronavirus

Convalescent plasma as a potential therapy for COVID-19

Plasma therapy against infectious pathogens, as of yesterday, today and tomorrow

Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein

Virological assessment of hospitalized patients with COVID-2019

The immunobiology of humanized Anti-IL6 receptor antibody: From basic research to breakthrough medicine

Tocilizumab: from the rheumatology practice to the fight against COVID-19, a virus infection with multiple faces

A decade of progress in tissue engineering

Tissue engineering and growth factors: updated evidence

Role of tissue engineering in COVID-19 and future viral outbreaks

In vitro models to study human lung development, disease and homeostasis

Tissue engineering: Still facing a long way ahead

Toward drug repurposing in epigenetics: olsalazine as a hypomethylating compound active in a cellular context

Drug treatment options for the 2019-new coronavirus (2019-nCoV)

Clinical characteristics and therapeutic procedure for four cases with 2019 novel coronavirus pneumonia receiving combined Chinese and Western medicine treatment

Genome-scale engineering for systems and synthetic biology

Breakthrough of the Year: CRISPR makes the cut

Genetic manipulation of genomes with rare-cutting endonucleases

Gene editing: not just for translation anymore

Tools for translation: non-viral materials for therapeutic mRNA delivery

Applications of CRISPR technologies in research and beyond

Applications of CRISPR/Cas9 for Gene Editing in Hereditary Movement Disorders

Development and applications of CRISPR-Cas9 for genome engineering

The new frontier of genome engineering with CRISPR-Cas9

Multiplex Genome Engineering Using CRISPR/Cas Systems

Cas9-mediated targeting of viral RNA in eukaryotic cells

CRISPR/Cas, the immune system of bacteria and archaea

DNA targeting specificity of RNA-guided Cas9 nucleases

Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems

Reflections on the early development of poxvirus vectors

C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector

Double-stranded RNA adenosine deaminases enhance expression of human immunodeficiency virus type 1 proteins

Evidence for host-dependent RNA editing in the transcriptome of SARS-CoV-2. Science Advances

The preferred nucleotide contexts of the AID/APOBEC cytidine deaminases have differential effects when mutating retrotransposon and virus sequences compared to host genes

Towards CRISPR/Cas crops -bringing together genomics and genome editing

CRISPR/Cas system mediates bacterial innate immune evasion and virulence

RNA targeting with CRISPR-Cas13

Genome editing in primary cells and in vivo using viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins

Coronavirus reverse genetics by targeted RNA recombination

Manipulation of the Coronavirus Genome Using Targeted RNA Recombination with Interspecies Chimeric Coronaviruses

High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases

Engineered Viruses as Genome Editing Devices

Recent advances in lentiviral vector development and applications

Ex vivo gene transfer and correction for cell-based therapies

Targeted gene correction minimally impacts whole-genome mutational load in human-disease-specific induced pluripotent stem cell clones

Pre-existing antiadenoin AAV gene therapy

Easy quantitative assessment of genome editing by sequence trace decomposition

Progress and prospects: zinc-finger nucleases as gene therapy agents

Efficient delivery of genomeediting proteins in vitro and in vivo

Nanotechnology on duty in medical applications

Nanopharmaceuticals: Innovative theranostics for the neurological disorders

New perspectives in biosensor technology

Engineering Nanomaterial Surfaces for Biomedical Applications

The role of nanotechnology in the treatment of viral infections

Interaction of silver nanoparticles with HIV-1

Nano-based approach to combat emerging viral (NIPAH virus) infection

Multiplexed colorimetric detection of Kaposi's sarcoma associated herpesvirus and Bartonella DNA using gold and silver nanoparticles

Inhibition of Herpes Simplex Virus Type 1 Infection by Silver Nanoparticles Capped with Mercaptoethane Sulfonate

Tannic Acid Modified Silver Nanoparticles Show Antiviral Activity in Herpes Simplex Virus Type 2 Infection

Applications of gold nanoparticles in virus detection

Colorimetric biosensors for point-of-care virus detections

Direct detection of unamplified hepatitis C virus RNA using unmodified gold nanoparticles

A highly selective and colorimetric assay J o u r n a l P r e -p r o o f Journal Pre-proof of lysine by molecular-driven gold nanorods assembly

Carbon nanotubes prolong the regulatory action of nerve growth factor on the endocannabinoid signaling

Application of carbon nanotubes as the carriers of the cannabinoid, 2-arachidonoylglycerol: Towards a novel treatment strategy in colitis

Nerve growth

Application of Carbon Nanotubes for Controlled Release of Growth Factors or Endocannabinoids: A Breakthrough in Biomedicine

Carbon nanotubes provide longer lasting gastroprotective effects for anandamide in stressinduced gastric ulcer in rat

Direct electron transfer of polyphenol oxidase on carbon nanotube surfaces: Application in Biosensing

Antibetanodavirus activity of isoprinosine and improved efficacy using carbon nanotubes based drug delivery system

Carbon nanotube-based nanocarrier loaded with ribavirin against grass carp reovirus

High aspect ratio nanomaterials enable delivery of functional genetic material without DNA integration in mature plants

Detection of Epstein-Barr virus infection in cancer by using highly specific nanoprobe based on dBSA capped CdTe quantum dots

Robust and highly sensitive fluorescence approach for point-of-care virus detection based on immunomagnetic separation

Nonviral Nanoparticles for CRISPR-Based Genome Editing: Is It Just a Simple Adaption of What Have Been Developed for Nucleic Acid Delivery?

High-throughput in vivo screen of functional mRNA delivery identifies nanoparticles for endothelial cell gene editing

Barcoded nanoparticles for high throughput in vivo discovery of targeted therapeutics

Ferulic acidloaded nanostructured lipid carriers: A promising nanoformulation against the ischemic neural injuries

Aerosol delivery of ferulic acid-loaded nanostructured lipid carriers: A promising treatment approach against the respiratory disorders

Therapeutic mRNA delivery to leukocytes

High-throughput in vivo screen of functional mRNA delivery identifies nanoparticles for endothelial cell gene editing

Lipid polymer hybrid nanomaterials for mRNA delivery

Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes

Sequence-engineered mRNA without chemical nucleoside modifications enables an effective protein therapy in large animals

Administration of nucleoside-modified mRNA encoding broadly neutralizing antibody protects humanized mice from HIV-1 challenge

Non-viral delivery of zinc finger nuclease mrna enables highly efficient in vivo genome editing of multiple therapeutic gene targets

Long-term storage of lipid-like nanoparticles for mRNA delivery

Microfluidic mixing: a general method for encapsulating macromolecules in lipid nanoparticle systems

Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA

Rapid discovery of potent siRNA-containing lipid nanoparticles enabled by controlled microfluidic formulation

Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads

Prospects of RNAi and microRNA-based therapies for hepatitis C

RNAi, a new therapeutic strategy against viral infection

Antiviral applications of RNAi for Coronavirus

siRNA-mediated antitumorgenesis for drug target validation and therapeutics

A systematic analysis of the silencing effects of an active siRNA at all single-nucleotide mismatched target sites

Progress in the use of RNA interference as a therapy for chronic hepatitis B virus infection

Knocking Down Barriers: Advances in siRNA Delivery

Advances in lipid nanoparticles for siRNA delivery

Cationic lipid nanosystems as carriers for nucleic acids

FDA approves patisiran to treat hereditary transthyretin amyloidosis

The Onpattro story and the clinical translation of nanomedicines containing nucleic acid-based drugs

Maximizing the Potency of siRNA Lipid Nanoparticles for Hepatic Gene Silencing In Vivo

Liver target delivery of small interfering RNA to the HCV gene by lactosylated cationic liposome

María Ángeles Solinís, and Ana del Pozo

Lipid Nanoparticles as Carriers for RNAi against Viral Infections: Current Status and Future Perspectives

Lipid nanoparticle siRNA treatment of Ebola virus Makona infected nonhuman primates

mRNA vaccine delivery using lipid nanoparticles

Nucleic acid vaccines: prospects for non-viral delivery of mRNA vaccines

Multi-antigenic human cytomegalovirus mRNA vaccines that elicit potent humoral and cell-mediated immunity

Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines

A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing

Nanoparticle-Based Delivery of CRISPR/Cas9 Genome-Editing Therapeutics

CRISPR genome engineering and viral gene delivery: a case of mutual attraction

Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR-Cas gene editing

Smart cancer nanomedicine: Strategic directions to improve translation and exploitation

Polymer-Lipid Nanoparticles for Systemic Delivery of mRNA to the Lungs

Direct cytosolic delivery of CRISPR/Cas9-polyamine-DNA polyplexes

Graphene oxide-mediated Cas9/sgRNA delivery for efficient genome editing

A biodegradable nanocapsule delivers a Cas9 ribonucleoprotein complex for in vivo genome editing

Antiviral applications of RNAi for Coronavirus

Prophylactic and therapeutic effects of small interfering RNA targeting SARS-coronavirus

Using siRNA in prophylactic and therapeutic regimens against SARS coronavirus in Rhesus macaque

Mechanisms of severe acute respiratory syndrome pathogenesis and innate immunomodulation. Microbiol

Small Interfering RNA Effectively Inhibits the Expression of SARS Coronavirus Membrane Gene at Two Novel Targeting Sites

Liver Disease Associated with Non-Hepatitis Viruses. Encyclopedia of Gastroenterology

Dendrimer-Based Lipid Nanoparticles Deliver Therapeutic FAH mRNA to Normalize Liver Function and Extend Survival in a Mouse Model of Hepatorenal Tyrosinemia Type I

Nanotechnology for organ-tunable gene editing

Artificial intelligence in nanotechnology

Computational modelling: moonlighting on the neuroscience and medicine

Ignoring the modeling approaches: Towards the shadowy paths in nanomedicine

Application of modelling and nanotechnology-based approaches: The emergence of breakthroughs in theranostics of central nervous system disorders

Molecular docking and structurebased drug design strategies

The structural model of Zika virus RNA-dependent RNA polymerase in complex with RNA for rational design of novel nucleotide inhibitors

Rapid Therapeutic Recommendations in the Context of a Global Public Health Crisis using Translational Bioinformatics Approaches: A proof-of-concept study using

Antiviral drug discovery: broadspectrum drugs from nature

Identification of contaminant sources in enclosed environments by inverse CFD modeling

Impact of scaling and body movement on contaminant transport in airliner cabins

COVID-19 Disease Map, building a computational repository of SARS-CoV-2 virus-host interaction Mechanisms

FBA reveals guanylate kinase as a potential target for antiviral therapies against SARS-CoV-2

Community-driven roadmap for integrated disease maps

Tools for visualization and analysis of molecular networks, pathways, and -omics data

A Biophysical Model of CRISPR/Cas9 Activity for Rational Design of Genome Editing and Gene Regulation

Medical nanorobot architecture based on nano bioelectronics

Nanorobotic Applications in Medicine: Current Proposals and Designs

Equipment and technology in robotics

Creation of nanorobots: Both state-of-the science and state-of-the-art

Nanorobot: A versatile tool in J o u r n a l P r e -p r o o f Journal Pre-proof nanomedicine

A remote-controlled generation of gold@polydopamine (core@ shell) nanoparticles via physical-chemical stimuli of polydopamine/gold composites

A computerassisted protocol for endovascular target interventions using a clinical MRI system for controlling untethered microdevices and future nanorobots

Micromachine-enabled capture and isolation of cancer cells in complex media

Nanorobotics ideas in nanomedicine

Interventional procedure based on nanorobots propelled and steered by flagellated magnetotactic bacteria for direct targeting of tumors in the human body

Nanobots: The future of medical treatments

A DNA origami nanorobot controlled by nucleic acid hybridization

A logic-gated nanorobot for targeted transport of molecular payloads

Bacterial flagella-based propulsion and on/off motion control of microscale objects

Computational studies of viral protein nano-actuators

Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin

A survey on wireless grid computing

Investigation into the future of RFID in biomedical applications

CRISPR-Cas9: from Genome Editing to Cancer Research

Nanotechnology based CRISPR/Cas9 system delivery for genome editing: Progress and prospect