Premium
Generation of apoptosis‐resistant HEK293 cells with CRISPR/Cas mediated quadruple gene knockout for improved protein and virus production
Author(s) -
Zhang Weifeng,
Xiao Dan,
Shan Linlin,
Zhao Junli,
Mao Qinwen,
Xia Haibin
Publication year - 2017
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26382
Subject(s) - apoptosis , hek 293 cells , cell culture , microbiology and biotechnology , crispr , biology , virus , biopharmaceutical , gene , chemistry , virology , genetics
Apoptosis has important functions during pathophysiologic processes. However, from a biopharmaceutical point of view, active apoptosis of host cells is undesirable during viral packaging or protein expression, because it decreases the efficiency of viral or protein production. Here we used the CRISPR/Cas technique to knock out four pro‐apoptotic genes, Caspase3, Caspase6, Caspase7 and AIF1 , in HEK293 cells, and successfully produced an apoptosis‐resistant cell line. Furthermore, this cell line showed higher expression levels of pro‐apoptotic proteins and higher packaging efficiency for the virus carrying these proteins than control HEK293 cells. This study not only produced an apoptosis‐resistant cell line that is useful in producing apoptosis‐inducing proteins or viruses expressing these proteins, but also provides a methodology to build other apoptosis‐resistant cell lines.