Construction and preliminary identification of MAVS or NFκB1 gene knockout in MDCK cells using CRISPR/Cas9
10.3760/cma.j.cn112866-20191115-00179
- VernacularTitle:基于CRISPR/Cas9技术的 MAVS和 NFκB1基因敲除MDCK细胞系的构建及初步鉴定
- Author:
Di TIAN
1
;
Yi LIU
;
Haili CHEN
;
Chunyi YANG
;
Wei WANG
;
Zhigang YI
;
Wanju ZHANG
Author Information
1. 上海市公共卫生临床中心 201508
- Keywords:
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR- associated 9 (Cas9);
Madin-Darby canine kidney cell (MDCK);
Mitochondrial ant
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(2):197-201
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct mitochondrial antiviral signaling ( MAVS) gene or Nuclear factor kappa B1 ( NFκB1) gene knockout Madin-Darby canine kidney (MDCK) cells, and identify the cell growth and proliferation characteristics of influenza virus (Flu) in these cells. Methods:MAVS or NFκB1 knockout MDCK cells were established using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) technique. Flu was inoculated into these cells, then the hemagglutination (HA) titers and TCID 50 were determined and compared with the original MDCK cells. Results:MDCK cells knocked out of MAVS (MDCK- MAVS-) or NFκB1 (MDCK- NFκB1 -) were obtained stably. The two cells were both adherent epithelioid cells as well as wild type MDCK cells. The HA titers showed no obvious difference among these cells after inoculating Flu. Nevertheless, the TCID 50 titers were respectively increased 9.5 times in MDCK- MAVS-cell culture and 10 times in MDCK- NFκB1 - cell culture, compared to the wild type MDCK cells. Conclusions:The infectivities of Flu were both increased in MDCK- MAVS-cells and MDCK- NFκB1 - cells, illustrating their potential in Flu culture.
