CRISPR-Cas9 mediated knockout of AnxA6 gene enhances influenza A virus replication in low-permissive HEK293FT cell line

CRISPR-Cas9 mediated knockout of AnxA6 gene enhances influenza A virus replication in low-permissive HEK293FT cell line Full article

Journal

GENE

Output data

Year: 2022, Volume: 809, Article number : 146024, Pages count : DOI: 10.1016/j.gene.2021.146024

Tags

Annexin A6; CRISPR-Cas9-mediated knockout; Genome editing; Influenza virus; Permissive cell line

Authors

Komissarov A. ^1,2 , Sergeeva M. ^1,2 , Zhuravlev E. ² , Medvedev S. ^2,3 , Malakhova A. ^2,3 , Andreeva E. ¹ , Shurygina A.-P. ¹ , Gorshkov A. ¹ , Timofeeva M. ^1,2 , Balakhonova E. ² , Grudinin M. ¹ , Zakian S. ^2,3 , Richter V. ² , Stepanov G. ²

Affiliations

1	Smorodintsev Research Institute of Influenza, Ministry of Health of Russian Federation, Prof. Popova 15/17, Saint Petersburg, 197376, Russian Federation
2	Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, Lavrentiev Avenue 8, Novosibirsk, 630090, Russian Federation
3	Federal Research Centre “Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences”, Lavrentiev Avenue 10, Novosibirsk, 630090, Russian Federation

Using cell cultures of human origin for the propagation of influenza virus is an attractive way to preserve its glycosylation profile and antigenic properties, which is essential in influenza surveillance and vaccine production. However, only few cell lines are highly permissive to influenza virus, and none of them are of human origin. The barrier might be associated with host restriction factors inhibiting influenza growth, such as AnxA6 protein counteracting the process of influenza virion packaging. In the presented work we explore the CRISPR-Cas9 mediated knockout of ANXA6 gene as a way to overcome the host restriction barrier and increase the susceptibility of human cell line to influenza infection. By CRISPR-Cas9 genome editing we modified HEK293FT cells and obtained several clones defective in the ANXA6 gene. The replication of the influenza A virus in original HEK293FT cells and the HEK293FT-ANXA6-/- mutant cells was compared in growth curve experiments. By combination of methods including TCID assay and flow cytometry we showed that accumulation of influenza A virus in the mutant HEK293FT-ANXA6-/- cells significantly exceeded the virus titer in the original HEK293FT cells. © 2021 Elsevier B.V.

Komissarov A. , Sergeeva M. , Zhuravlev E. , Medvedev S. , Malakhova A. , Andreeva E. , Shurygina A.-P. , Gorshkov A. , Timofeeva M. , Balakhonova E. , Grudinin M. , Zakian S. , Richter V. , Stepanov G.
CRISPR-Cas9 mediated knockout of AnxA6 gene enhances influenza A virus replication in low-permissive HEK293FT cell line
GENE. 2022. V.809. 146024 . DOI: 10.1016/j.gene.2021.146024 WOS Scopus РИНЦ OpenAlex

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