Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2539-2549 |
| Number of pages | 11 |
| Journal | Biotechnology and Bioengineering |
| Volume | 114 |
| Issue number | 11 |
| DOIs | |
| State | Published - Nov 2017 |
Funding
National Natural Science Foundation of China, Grant numbers: 31601086, 81471772; Natural Science Foundation of Shaanxi Province, Grant number: 2014JZ005; China Postdoctoral Science Foundation, Grant number: 2015M570809; Fundamental Research Funds for the Central Universities, Grant number: GK201603113 This work was supported by the Fundamental Research Funds for the Central Universities (GK201603113), the China Postdoctoral Science Foundation (2015M570809), research grants to H.X. and W.Z. from the National Natural Science Foundation of China (No. 81471772 and No.31601086), and research grants to H.X from the Natural Science Foundation of Shaanxi Province, China (No. 2014JZ005).
Keywords
- CRISPR/Cas
- adenovirus
- apoptosis
- gene therapy
- genome editing
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Bioengineering
- Biotechnology