Abstract
Coronavirus disease 2019 (COVID-19) is a new respiratory illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and now spreads globally. Currently, therapeutics and effective treatment options remain scarce and there is no proven drug to treat COVID-19. Targeting the positive-sense RNA genome and viral mRNAs of SARS-CoV-2 to simultaneously degrade viral genome templates for replication and viral mRNAs for essential gene expression would be a strategy to completely realize virus elimination. Type VI CRISPR enzymes Cas13 have recently been identified as programmable RNA-guided, RNA-targeting Cas proteins with nuclease activity that allows for RNA cleavage and degradation. The precise viral RNA detection and antiviral application of the CRISPR/Cas13 system depend on high-efficient and minimal off-target crRNAs. Although a computer-based algorithm has been applied for the design of crRNAs targeting SRAS-CoV-2, the experimental screening system to identify optimal crRNA is not available. We develop a one-step experimental screening system to identify high-efficient crRNAs with minimal off-target effects for CRISPR/Cas13-based SARS-CoV-2 elimination. This platform provides the foundation for CRISPR/Cas13-based diagnostics and therapeutics for COVID-19. This platform is versatile and could also be applied for crRNAs screening for other RNA viruses.
Keywords: ACE2, Angiotensin-converting enzyme 2; COVID-19; COVID-19, Coronavirus disease 2019; CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR/Cas13; SARS-CoV-2; SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus 2; scRNA-seq; scRNA-seq, Single-cell RNA-seq.
【저자키워드】 COVID-19, SARS-CoV-2, COVID-19, Coronavirus disease 2019, SARS-CoV-2, Severe acute respiratory syndrome coronavirus 2, ACE2, angiotensin-converting enzyme 2, scRNA-seq, CRISPR/Cas13, CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats, scRNA-seq, Single-cell RNA-seq., 【초록키워드】 Treatment, coronavirus disease, severe acute respiratory syndrome coronavirus 2, Coronavirus disease 2019, ACE2, coronavirus, Gene Expression, Antiviral, Therapeutics, Respiratory illness, severe acute respiratory syndrome Coronavirus, angiotensin-converting enzyme 2, diagnostics, RNA, Replication, Spread, Protein, CRISPR, RNA-Seq, RNA viruses, Algorithm, single-cell RNA-seq, cleavage, target, Viral RNA, Degradation, respiratory, disease, scRNA-seq, platform, SRAS-CoV-2, Angiotensin-converting enzyme, angiotensin, viral genome, Nuclease, clustered regularly interspaced short palindromic repeats, acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, enzyme, foundation, treat, RNA genome, targeting, virus elimination, viral mRNAs, Effect, effective, positive-sense, identify, develop, caused, applied, provide, Type, viral mRNA, degrade, Palindromic, 【제목키워드】 Treatment, SARS-CoV-2 virus, platform, COVID-19 patient,