title: Inhibiting cyprinid herpesvirus-3 replication with CRISPR/Cas9.
abstract: OBJECTIVE
  The potential of CRISPR/Cas9 gene editing to repress CyHV-3 was tested in vitro.
  
  
  RESULTS
  By targeting two basic target genes necessary for the early transcription of CyHV-3, we show that virus transcription and particle release were significantly decreased by CRISPR/Cas9, as measured by quantitative real-time PCR and virus titration experiments, respectively.
  
  
  CONCLUSIONS
  (A) The effectiveness is confirmed of the CRISPR/Cas9 system at repressing exogenous genes, including large viral genomic DNA, by introducing site-specific mutations in vitro. (B) The CyHV-3 virus replicates poorly in Cas9-positive cells. (C) The inhibition of thymidine kinase alone cannot block viral particle release.
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin, China.|College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Yicheng
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Tiedong
authors.initials: T
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin, China.
authors.forename: Ze
authors.initials: Z
authors.lastname: Yu
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin, China.
authors.forename: Heming
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Animal Sciences, Jilin University, Changchun, China.|College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Liu
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin, China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Wu
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin, China. chunboteng@nefu.edu.cn.
authors.forename: Chun-Bo
authors.initials: CB
authors.lastname: Teng
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D002347:Carps / Q000821:virology*; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D006564:Herpesviridae / Q000502:physiology*; D014779:Virus Replication
chemical_list: D004279:DNA, Viral
keywords: CRISPR/Cas9; Cyprinid herpesvirus-3; DNA polymerase; Koi herpesvirus; Thymidine kinase