期刊
JOURNAL OF VIROLOGY
卷 91, 期 8, 页码 -出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.02465-16
关键词
Bombyx mori nucleopolyhedrovirus; CRISPR/Cas9; transgenic silkworm; antiviral therapy
类别
资金
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDB11010600]
- National Basic Research Program of China [2015CB755703]
- National Science Foundation of China [31420103918, 31530072]
- Chinese Academy of Sciences [KJZD-EW-L12-02]
We developed a novel antiviral strategy by combining transposonbased transgenesis and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system for the direct cleavage of Bombyx mori nucleopolyhedrovirus (BmNPV) genome DNA to promote virus clearance in silkworms. We demonstrate that transgenic silkworms constitutively expressing Cas9 and guide RNAs targeting the BmNPV immediate early-1 (ie-1) and me53 genes effectively induce target-specific cleavage and subsequent mutagenesis, especially large (similar to 7-kbp) segment deletions in BmNPV genomes, and thus exhibit robust suppression of BmNPV proliferation. Transgenic animals exhibited higher and inheritable resistance to BmNPV infection than wild-type animals. Our approach will not only contribute to modern sericulture but also shed light on future antiviral therapy. IMPORTANCE Pathogen genome targeting has shown its potential in antiviral research. However, transgenic CRISPR/Cas9 system-mediated viral genome targeting has not been reported as an antiviral strategy in a natural animal host of a virus. Our data provide an effective approach against BmNPV infection in a real-world biological system and demonstrate the potential of transgenic CRISPR/Cas9 systems in antiviral research in other species.
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