Key result
CRISPR/Cas13d reduces SARS-CoV-2 mRNA and protein expression in vitro with ~80% editing efficiency.
Why the study?
There are currently very limited effective methods to prevent and control SARS-CoV-2 infection, and an efficient and specific mechanism to prevent COVID-19 is urgently needed.
Answers are grounded in this paper and cite their sources.
The CRISPR/Cas13d system demonstrates potential as a targeted antiviral strategy by effectively editing SARS-CoV-2 spike and nucleocapsid RNA in vitro.
Background As a new type of virus that seriously threatens public health worldwide, there are currently very limited effective methods of prevention and control for severe acute respiratory syndrome (SARS)-CoV-2. Therefore, people urgently need an efficient and specific mechanism to prevent COVID-19. CRISPR/Cas13 is a natural defense system in bacteria that has been successfully applied for the prevention and treatment of many viruses, including SARS-CoV-2. How to efficiently and specifically destroy SARS-CoV-2 RNA and avoid virus escape caused by base mutations is still an urgent problem. Methods By comparing and analyzing existing sequences in NCBI, highly conserved segments of protein-coding sequences were obtained to construct a highly specific crRNA plasmid. The A549 cell line stably expressing Cas13d was constructed by a lentivirus system. The cleavage efficiency of Cas13d for the target protein RNA of SARS-CoV-2 was confirmed by qPCR, WB, flow cytometry and immunofluorescence. Results This study focused on the spike (S) glycoprotein and nucleocapsid (N) protein. We found that the CRISPR/Cas13d system was able to effectively edit the SARS-CoV-2 genes at the mRNA or protein level. However, the results also showed that the combination of multiple crRNA could not effectively further improve the editing efficiency. Conclusions The CRISPR/Cas13 system was able to edit spike and nucleocapsid viral RNA to resist SARS-CoV-2 infection. This study provides strong theoretical support for the prevention and treatment of COVID-19.
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Wang et al. (2023) studied SARS-CoV-2. CRISPR/Cas13d system with specific crRNAs vs. Mock crRNA was evaluated on Reduction in viral mRNA and protein expression. The CRISPR/Cas13d system effectively reduced SARS-CoV-2 spike and nucleocapsid mRNA and protein expression in vitro, with S1crRNA achieving over 80% editing efficiency at the mRNA level.
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