%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/