Progress in crispr/Cas13-mediated suppression of influenza A and SARS-CoV-2 virus infection in in vitro and in vivo models

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Abstract

The worldwide number of deaths from complications caused by severe influenza and COVID-19 is about 1 million cases annually. The development of effective antiviral therapy strategies for the disease treatment is one of the most important tasks. The use of the CRISPR/Cas13 system, which specifically degrades viral RNA and significantly reduces the titer of the virus, can be a solution of this problem. Despite the recent discovery, Cas13 nucleases have already shown their high efficiency in suppressing viral transcripts in cell cultures. Recent advances in mRNA technology and improvements in non-viral delivery systems have made it possible to effectively use CRISPR/Cas13 in animal models as well. In this review, we have analyzed the experimental in vitro and in vivo studies on the use of CRISPR/Cas13 systems as antiviral agent in cell cultures and animals and discussed the main directions for improving the CRISPR/Cas13 system. These data allow us to understand the prospects and limitations of further use of CRISPR/Cas13 in the treatment of viral diseases.

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About the authors

A. A. Kazakova

Sirius University of Science and Technology

Email: reshetnikov.vv@talantiuspeh.ru
Russian Federation, 354340 Federal Territory “Sirius”

E. I. Leonova

Saint-Petersburg State University

Email: e.leonova@spbu.ru
Russian Federation, 199034 Saint-Peterburg

J. V. Sopova

Saint-Petersburg State University

Email: e.leonova@spbu.ru
Russian Federation, 199034 Saint-Peterburg

A. V. Chirinskaite

Saint-Petersburg State University

Email: e.leonova@spbu.ru
Russian Federation, 199034 Saint-Peterburg

E. S. Minskaya

Sirius University of Science and Technology

Email: reshetnikov.vv@talantiuspeh.ru
Russian Federation, 354340 Federal Territory “Sirius”

I. S. Kukushkin

Sirius University of Science and Technology

Email: reshetnikov.vv@talantiuspeh.ru
Russian Federation, 354340 Federal Territory “Sirius”

R. A. Ivanov

Sirius University of Science and Technology

Email: reshetnikov.vv@talantiuspeh.ru
Russian Federation, 354340 Federal Territory “Sirius”

V. V. Reshetnikov

Sirius University of Science and Technology; Institute of Cytology and Genetics, Siberian Branchn Academy of Sciences

Author for correspondence.
Email: reshetnikov.vv@talantiuspeh.ru
Russian Federation, 354340 Federal Territory “Sirius”; 630090 Novosibirsk

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2. Fig. 1. Structural and non-structural proteins encoded in the SARS-CoV genome. Non-structural proteins nsp (1–16) play important roles in viral replication and are encoded in ORF1ab. Polyproteins PP1a and PP1ab are cleaved by viral proteases PLpro (papain-like protease) and 3CLpro (3C-like major protease) to active viral proteins. Structural and accessory proteins are encoded in the 3′-terminal part of the genome

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3. Fig. 2. Schematic representation of the influenza A virus genome. a – Replication of influenza virus and other RNA viruses with (−) RNA genome. The ribonucleic acid complex (RNP), consisting of (−) genomic RNA in a complex with NP, cannot be translated and produce viral proteins. The viral polymerase RdRp, interacting with the RNP, first transcribes (+) mRNA, from which the viral proteins necessary for viral replication are translated, which then participate in the replication of new genomic (−) RNA. b – 8 RNA segments encode viral proteins. Cas13 targets are highlighted in red

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4. Fig. 3. Diversity of Cas13 enzymes. a – Discovered and characterized enzymes of the Cas13 family with the degree of phylogenetic relationship (based on the materials of the following articles: [37–39]). b – Enzymes of the Cas13 family used for inactivation of viruses in cell cultures and in vivo (based on the materials of the following articles: [3, 37, 38, 41, 42])

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