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• W2540875078 abstract "The CRISPR-associated endonuclease Cas9 can edit particular genomic loci directed by a single guide RNA (gRNA). The CRISPR/Cas9 system has been successfully employed for editing genomes of various organisms. Here we describe a protocol for editing the vaccinia virus (VV) genome in the cytoplasm of VV-infected CV-1 cells using the RNA-guided Cas9. RNA-guided Cas9 induces double-stranded DNA breaks facilitating homologous recombination efficiently and specifically in the targeted site of VV and a transgene can be incorporated into these sites by homologous recombination. By using a site-specific homologous vector with transgene(s), a N1L gene-deleted VV with the red fluorescence protein (RFP) gene incorporated in this region was generated with a successful recombination efficiency 10 times greater than that obtained from the conventional homologous recombination method. This protocol demonstrates successful use of RNA-guided Cas9 system to generate mutant VVs with enhanced efficiency." @default.
• W2540875078 created "2016-11-04" @default.
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• W2540875078 date "2016-10-30" @default.
• W2540875078 modified "2023-10-16" @default.
• W2540875078 title "A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors" @default.
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• W2540875078 doi "https://doi.org/10.3791/54171" @default.
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