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

作     者：Lin-zhu Ren Zhi-yuan Peng Ting Ouyang Xiao-hui Liu Xin-rong Chen Li Ye Jun-wen Fan Hong-sheng Ouyang Da-xin Pang Jie-ying Bai 

作者机构：Jilin Provincial Key Laboratory of Animal Embryo Engineering College of Animal Sciences Jilin University The Laboratory Animal Center The Academic of Military Medical Sciences 

出 版 物：《Animal Models and Experimental Medicine》 (动物模型与实验医学（英文）)

年 卷 期：2018年第1卷第1期

页      面：74-77页

学科分类：10[医学] 

基　　金：financially supported by the National Key Research and Development Program of China(No.2017YFD0500103) the Beijing Natural Science Foundation(No.5152023) the National Natural Science Foundation of China(No.31772747 and31272385) the Jilin Province Science and Technology Development Projects(20150204077NY) the Graduate Innovation Fund of Jilin University the Program for Chang jiang Scholars the University Innovative Research Team(No.IRT1248) 

主　　题：CRISPR/Cas9 pseudorabies virus(PRV) single-guide RNA(sgRNA) UL30 protein 

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 *** 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.