%0 Journal Article
%T Subculturing cells have no effect on CRISPR/Cas9©\mediated cleavage of UL30 gene in pseudorabies virus
%A Da©\xin Pang
%A Hong©\sheng Ouyang
%A Jun©\wen Fan
%A Li Ye
%A Lin©\zhu Ren
%A Ting Ouyang
%A Xiao©\hui Liu
%A Xin©\rong Chen
%A Zhi©\yuan Peng
%J Archive of "Animal Models and Experimental Medicine".
%D 2018
%R 10.1002/ame2.12006
%X 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
%K CRISPR/Cas9
%K pseudorabies virus (PRV)
%K single©\guide RNA (sgRNA)
%K UL30 protein
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6357676/