Knockdown the expression of ku70 and lig4 in HEK293T cells by CRISPR/Cas13 system.

Haoqiang Wang; Guoling LI; Guangyan Huang; Zicong LI; Enqin Zheng; Zheng XU; Huaqiang Yang; Zhenfang WU; Xianwei Zhang; Dewu Liu

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Knockdown the expression of ku70 and lig4 in HEK293T cells by CRISPR/Cas13 system.

Author: Haoqiang WANG ¹ ; Guoling LI ¹ ; Guangyan HUANG ¹ ; Zicong LI ¹ ; Enqin ZHENG ¹ ; Zheng XU ¹ ; Huaqiang YANG ¹ ; Zhenfang WU ¹ ; Xianwei ZHANG ¹ ; Dewu LIU ¹
Author Information

1. National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou 510642, Guangdong, China.
Publication Type:Journal Article
Keywords: CRISPR/Cas13; gene knockdown; ku70; lig4
MeSH: CRISPR-Cas Systems; DNA Ligase ATP; genetics; Gene Expression Regulation; genetics; Gene Knockdown Techniques; HEK293 Cells; Humans; Ku Autoantigen; genetics
From: Chinese Journal of Biotechnology 2020;36(7):1414-1421
CountryChina
Language:Chinese
Abstract: Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.