title: Establishment of a highly efficient virus-inducible CRISPR/Cas9 system in insect cells.
abstract: Although current antiviral strategies can inhibit baculovirus infection and decrease viral DNA replication to a certain extent, novel tools are required for specific and accurate elimination of baculovirus genomes from infected insects. Using the newly developed clustered regularly interspaced short palindromic repeats/associated protein 9 nuclease (CRISPR/Cas9) technology, we disrupted a viral genome in infected insect cells in vitro as a defense against viral infection. We optimized the CRISPR/Cas9 system to edit foreign and viral genome in insect cells. Using Bombyx mori nucleopolyhedrovirus (BmNPV) as a model, we found that the CRISPR/Cas9 system was capable of cleaving the replication key factor ie-1 in BmNPV thus effectively inhibiting virus proliferation. Furthermore, we constructed a virus-inducible CRISPR/Cas9 editing system, which minimized the probability of off-target effects and was rapidly activated after viral infection. This is the first report describing the application of the CRISPR/Cas9 system in insect antiviral research. Establishment of a highly efficient virus-inducible CRISPR/Cas9 system in insect cells provides insights to produce virus-resistant transgenic strains for future.
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Zhan-Qi
authors.initials: ZQ
authors.lastname: Dong
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China; Institute of Life Sciences, Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Ming-Ya
authors.initials: MY
authors.lastname: Cao
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Fei-Fan
authors.initials: FF
authors.lastname: Dong
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Ya-Ming
authors.initials: YM
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing, 400716, China. Electronic address: lucheng@swu.edu.cn.
authors.forename: Cheng
authors.initials: C
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing, 400716, China. Electronic address: pmh047@126.com.
authors.forename: Min-Hui
authors.initials: MH
authors.lastname: Pan
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D012831:Bombyx; D064130:CRISPR-Associated Proteins; D064113:CRISPR-Cas Systems; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D000072669:Gene Editing; D015870:Gene Expression; D023061:Gene Order; D018390:Gene Targeting; D017930:Genes, Reporter; D005814:Genes, Viral; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral; D007313:Insecta; D000094704:RNA, Guide, CRISPR-Cas Systems
chemical_list: D064130:CRISPR-Associated Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems
keywords: Antiviral; BmNPV; Bombyx mori; CRISPR/Cas9; Virus-inducible CRISPR/Cas9 system