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Generation of an Oocyte-Specific Cas9 Transgenic Mouse for Genome Editing.

ABSTRACT: The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage embryos. Through comprehensive analysis, we also show allele complexity and off-target mutagenesis induced by this strategy is obviously lower than Cas9 mRNA/sgRNA injection. Thus, injection of sgRNAs into oocyte-specific Cas9 transgenic mouse embryo provides a convenient, efficient and reliable approach for mouse genome editing.

SUBMITTER: Zhang L

PROVIDER: S-EPMC4847922 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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Generation of an Oocyte-Specific Cas9 Transgenic Mouse for Genome Editing.

Zhang Linlin L Zhou Jiankui J Han Jinxiong J Hu Bian B Hou Ningning N Shi Yun Y Huang Xingxu X Lou Xin X

PloS one 20160427 4

The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage emb ...[more]

PMID: 27119535

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