中国比较医学杂志2016，Vol.26Issue(12)：1-4,4.DOI:10.3969/j.issn.1671-7856.2016.12.001
利用 CRISPR／Cas9技术构建 miRNA－29 b1基因敲除小鼠
Construction of miRNA-29b1 knockout mice based on CRISPR/Cas9 technology
摘要
Abstract
Objective To construct miRNA-29b1 gene knockout mice based on CRISPR/Cas9 technology. Methods To design and synthesize sgRNA according to the miRNA-29b1 sequence in Genbank .sgRNA and Cas9 were transcribed to RNA in vitro, these RNA were then microinjected into zygotes of C 57BL/6 mice.After mouse birth, the genome DNA was extracted and sequenced to identify its genotype; meanwhile , real-time PCR was used to assay the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney of mutated mice .Result A 20 bp sgRNA targeted on miRNA-29b1 was synthesized and transcribed to RNA with Cas 9.After microinjection, miRNA-29b1 gene-mutated mice were obtained.The sequencing results showed that there were two types of genotype for the mutated mice , one was 10 bp deletion, and another was 23 bp deletion accompanied with a 3 bp insertion.Compared with the wild-type mice, the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney was reduced significantly .Conclusions miRNA-29b1 gene knockout mice are constructed successfully by using CRISPR /Cas9 technology.关键词
CRISPR/Cas9/miRNA-29b1/基因敲除小鼠Key words
CRISPR/Cas9/miRNA-29b1/Gene knockout mice分类
医药卫生引用本文复制引用
赵勇,张海,师长宏,赵亚,辛智倩,刘佩娟,张彩勤,白冰,白杰英,王华..利用 CRISPR／Cas9技术构建 miRNA－29 b1基因敲除小鼠[J].中国比较医学杂志,2016,26(12):1-4,4.基金项目
国家自然科学基金优秀青年基金项目（NO．81522009）；国家自然科学基金面上项目（NO．8137257，81272385）；军队重点项目（NO．BWS14J058）。 （）
