Subculturing cells have no effect on CRISPR/Cas9‐mediated cleavage of UL30 gene in pseudorabies virus

Animal Models and Experimental Medicine

Li Ye¹  zhu Ren¹  Ting Ouyang¹  rong Chen¹  yuan Peng¹  Lin‐¹  Zhi‐¹  Xin‐²  hui Liu²  Xiao‐² 

[1] Jilin Provincial Key Laboratory of Animal Embryo Engineering College of Animal Sciences Jilin University Changchun Jilin China;The Laboratory Animal Center The Academic of Military Medical Sciences Beijing China

关键词: CRISPR/Cas9;    pseudorabies virus (PRV);    single‐;    guide RNA (sgRNA);    UL30 protein;   

DOI  :  10.1002/ame2.12006

学科分类：医学（综合）

来源: Wiley

【 摘 要 】

CRISPR/Cas9‐mediated genome editing can inhibit virus infection by targeting the conserved regions of the viral genomic DNA. Unexpectedly, we found previously that pseudorabies virus (PRV) could escape from CRISPR/Cas9‐mediated inhibition. In order to elucidate whether the escape of PRV from Cas9‐mediated inhibition was due to cell deficiencies, such as genetic instability of sgRNA or Cas9 protein, the positive cells were passaged ten times, and PRV infection in the sgRNA‐expressing cells was evaluated in the present study. The results showed that subculturing cells has no effect on Cas9‐mediated cleavage of PRV. Different passages of PX459‐PRV cells can stably express sgRNA to facilitate Cas9/sgRNA cleavage on the UL30 gene of PRV, resulting in a pronounced inhibition of PRV infection. Studies to elucidate the mechanism of PRV escape are currently in progress.

【 授权许可】

CC BY