Establishment of a highly efficient virus-inducible CRISPR/Cas9 system in insect cells

• Published in: Antiviral research Vol. 130; pp. 50 - 57
• Main Authors: Dong, Zhan-Qi, Chen, Ting-Ting, Zhang, Jun, Hu, Nan, Cao, Ming-Ya, Dong, Fei-Fan, Jiang, Ya-Ming, Chen, Peng, Lu, Cheng, Pan, Min-Hui
• Format: Journal Article
• Language: English
• Published: Netherlands 01.06.2016
• Subjects: Animals; Baculoviridae - genetics; Baculovirus; Base Sequence; Bombyx; Bombyx mori NPV; Cell Line; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR-Associated Proteins; CRISPR-Cas Systems; Gene Editing; Gene Expression; Gene Order; Gene Targeting; Genes, Reporter; Genes, Viral; Genetic Vectors - genetics; Genome, Viral; Insecta; RNA, Guide, CRISPR-Cas Systems; Virus-inducible CRISPR/Cas9 system; Antiviral; Bombyx mori; CRISPR/Cas9; BmNPV
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/j.antiviral.2016.03.009

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.