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Review
. 2021 Nov 11;13(11):2258.
doi: 10.3390/v13112258.

CRISPR Screening: Molecular Tools for Studying Virus-Host Interactions

Vladimir Chulanov  1   2   3 , Anastasiya Kostyusheva  1 , Sergey Brezgin  1   2 , Natalia Ponomareva  1   4 , Vladimir Gegechkori  4 , Elena Volchkova  3 , Nikolay Pimenov  1 , Dmitry Kostyushev  1   2   3
Affiliations
Review

CRISPR Screening: Molecular Tools for Studying Virus-Host Interactions

Vladimir Chulanov et al. Viruses. .

Abstract

CRISPR/Cas is a powerful tool for studying the role of genes in viral infections. The invention of CRISPR screening technologies has made it possible to untangle complex interactions between the host and viral agents. Moreover, whole-genome and pathway-specific CRISPR screens have facilitated identification of novel drug candidates for treating viral infections. In this review, we highlight recent developments in the fields of CRISPR/Cas with a focus on the use of CRISPR screens for studying viral infections and identifying new candidate genes to aid development of antivirals.

Keywords: COVID-19; Cas12; Cas9; HAV; HIV; SARS-CoV-2; ZIKV; hepatitis.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
General workflow of CRISPR screens. CRISPR screening protocols include 6 main steps: (1) in silico design of gRNA libraries; (2) cloning and validation of gRNA library; (3) packaging gRNAs into lentiviruses and transduction of lentiviruses into Cas-expressing cells; (4) alteration of genes in experimental conditions; (5) isolation of nucleic acids; and (6) deep sequencing of barcoded gRNAs and data analysis. Note: if using pre-made gRNA libraries, the first two steps of the protocol are omitted.
Figure 2
Figure 2
Positive and negative selection of cells for CRISPR screening. Cas-producing cells are transduced with the lentiviral gRNA library. Next, cells are infected with a cytolytic virus and then undergo positive selection (resistant cells are enriched while susceptible cells die) or negative selection (cells with altered antiviral genes die).
See this image and copyright information in PMC

References

    1. Brezgin S., Kostyusheva A., Kostyushev D., Chulanov V. Dead Cas Systems: Types, Principles, and Applications. Int. J. Mol. Sci. 2019;20:6041. doi: 10.3390/ijms20236041. - DOI - PMC - PubMed
    1. Brezgin S., Kostyusheva A., Bayurova E., Volchkova E., Gegechkori V., Gordeychuk I., Glebe D., Kostyushev D., Chulanov V. Immunity and Viral Infections: Modulating Antiviral Response via CRISPR–Cas Systems. Viruses. 2021;13:1373. doi: 10.3390/v13071373. - DOI - PMC - PubMed
    1. Parnas O., Jovanovic M., Eisenhaure T.M., Herbst R.H., Dixit A., Ye C.J., Przybylski D., Platt R.J., Tirosh I., Sanjana E.N., et al. A genome-wide CRISPR screen in primary immune cells to dissect regulatory networks. Cell. 2015;162:675–686. doi: 10.1016/j.cell.2015.06.059. - DOI - PMC - PubMed
    1. Zetsche B., Gootenberg J.S., Abudayyeh O.O., Slaymaker I.M., Makarova K.S., Essletzbichler P., Volz S.E., Joung J., van der Oost J., Regev A., et al. Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015;163:759–771. doi: 10.1016/j.cell.2015.09.038. - DOI - PMC - PubMed
    1. Wang X., Li J., Wang Y., Yang B., Wei J., Wu J., Wang R., Huang X., Chen J., Yang L. Efficient base editing in methylated regions with a human APOBEC3A-Cas9 fusion. Nat. Biotechnol. 2018;36:946. doi: 10.1038/nbt.4198. - DOI - PubMed

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