DOI: 10.1002/ame2.12006
Subculturing cells have no effect on CRISPR/Cas9-mediated cleavage of UL30 gene in pseudorabies virus
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
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.
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资助基金：This work was financially supported by the National Key Research and Development Program of China(2017YFD0500103)the Beijing Natural Science Foundation(5152023)the National Natural Science Foundation of China(31772747 and 31272385)the Jilin Province Science and Technology Development Projects(20150204077NY)the Graduate Innovation Fund of Jilin University, the Program for Chang jiang Scholars and the University Innovative Research Team(IRT1248)
论文发表日期：2018-01-01
在线出版日期：2025-08-15（本平台首次上网日期，不代表文献的发表时间）
页数：4( 74-77 )
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