A rapid and efficient platform for antiviral crRNA screening using CRISPR-Cas13a-based nucleic acid detection

Frontiers in Immunology

Immunology

Xue Dong¹  Hao Li¹  Yansong Sun¹  Wenyanbo Yi¹  Lan Yang¹  Yingjie Song¹  Yao Han¹  Mengwei Niu¹  Youcui Zhang² 

[1] State Key Laboratory of Pathogens and Biosafety, Beijing Institute of Microbiology and Epidemiology, Beijing, China;State Key Laboratory of Pathogens and Biosafety, Beijing Institute of Microbiology and Epidemiology, Beijing, China;College of Life Sciences, Fujian Agriculture and Forestry University, Fujian, China;

关键词: CRISPR/Cas13a;    crRNA screening;    antiviral;    nucleic acid detection;    influenza A(H1N1);   

DOI  :  10.3389/fimmu.2023.1116230

 received in 2022-12-05, accepted in 2023-04-24,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionRapid and high-throughput screening of antiviral clustered regularly interspaced short palindromic repeat (CRISPR) RNAs (crRNAs) is urgently required for the CRISPR-Cas13a antiviral system. Based on the same principle, we established an efficient screening platform for antiviral crRNA through CRISPR-Cas13a nucleic acid detection. MethodIn this study, crRNAs targeting PA, PB1, NP, and PB2 of the influenza A virus (H1N1) were screened using CRISPR-Cas13a nucleic acid detection, and their antiviral effects were confirmed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The RNA secondary structures were predicted by bioinformatics methods.ResultsThe results showed that crRNAs screened by CRISPR-Cas13a nucleic acid detection could effectively inhibit viral RNA in mammalian cells. Besides, we found that this platform for antiviral crRNA screening was more accurate than RNA secondary structure prediction. In addition, we validated the feasibility of the platform by screening crRNAs targeting NS of the influenza A virus (H1N1).DiscussionThis study provides a new approach for screening antiviral crRNAs and contributes to the rapid advancement of the CRISPR-Cas13a antiviral system.

【 授权许可】

Unknown   
Copyright © 2023 Yang, Zhang, Yi, Dong, Niu, Song, Han, Li and Sun