【摘 要】
：
The type Ⅱ bacterial CRISPR/Cas system is a very valuable genome-engineering tool, which has been widely used in genome editing of variety of organisms.Previously, we generated the floxed alleles in r
【机 构】
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Key Laboratory of Human Disease Comparative Medicine,Ministry of Health,Institute of Laboratory Anim
【出 处】
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2014第十一届中国实验动物科学年会
论文部分内容阅读
The type Ⅱ bacterial CRISPR/Cas system is a very valuable genome-engineering tool, which has been widely used in genome editing of variety of organisms.Previously, we generated the floxed alleles in rats by CRISPR/Cas9.Here, we successfully use two-cut strategy with one circular vector, which contains the exogenous cDNAs with homology arm regions, in generating knockin rats at Trdmt1, Nestin and Cck loci.The efficiency of CRISPR/Cas9-mediated knockin was up to 54%.Furthermore, by crossing Nestin-Cre rat with the Dnmt3b floxed rat and Cck-Cre with Dnmt1 floxed rat, we detected the Cre/loxP-mediated recombination in the F1 generation of rats.We also show that the knockin alleles were germline transmitted.These results firstly provided a simple and flexible engineering strategy for establishment of knockin rats.
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