Mar 17, 2014

CRISPR/Cpf1 mediated genome editing enhances Bombyx mori resistance to BmNPV

BioRxiv : the Preprint Server for Biology
Rowena BallMinhui Pan

Abstract

CRISPR/Cas12a (Cpf1) is a single RNA-guided endonuclease that provides new opportunities for targeted genome engineering through the CRISPR/Cas9 system. Only AsCpf1 have been developed for insect genome editing, and the novel Cas12a orthologs nucleases and editing efficiency require more study in insect. We compared three Cas12a orthologs nucleases, AsCpf1, FnCpf1, and LbCpf1, for their editing efficiencies and antiviral abilities in vitro. The three Cpf1 efficiently edited the BmNPV genome and inhibited BmNPV replication in BmN-SWU1 cells. The antiviral ability of the FnCpf1 system was more efficient than the SpCas9 system after infection by BmNPV. We created FnCpf1×gIE1 and SpCas9×sgIE1 transgenic hybrid lines and evaluated the gene editing efficiency of different systems at the same target site. We improved the antiviral ability using the FnCpf1 system in transgenic silkworm. This study demonstrated use of the CRISPR/Cpf1 system to achieve high editing efficiencies in the silkworm, and illustrates the use of this technology for increasing disease resistance.

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Mentioned in this Paper

Solocyte
Hydrogen Peroxide
RNA, Double-Stranded
Prebiotics
Virus Replication
Oscillatory Muscle Contraction
Complementary RNA
Species
Surface
Brain Waves

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