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Sci.Rep:上海交通大学陈学进研究组发表基因敲除克隆兔的研究成果

摘要 : 11月2日,Nature出版集团旗下期刊《Scientific Reports》在线发表上海交通大学医学院实验动物科学部陈学进研究小组的最新研究成果.

 11月2日,Nature出版集团旗下期刊《Scientific Reports》在线发表上海交通大学医学院实验动物科学部陈学进研究小组的最新研究成果,研究论文题为“Generation of hypoxanthinephosphoribosyltransferase geneknockout rabbits by homologousrecombination and gene trappingthrough somatic cell nucleartransfer”(利用核移植及同源重组和基因捕获技术获得HPRT基因敲除兔)。陈学进研究组博士生尹明茹和姜伟华并列为论文第一作者,李善刚副研究员与陈学进研究员为共同通讯作者。

次黄嘌呤鸟嘌呤磷酸核糖转移酶(Hypoxanthine-guanine phosphoribosyltransferase,HPRT)在嘌呤核苷酸的补救合成过程中起重要作用,在脑组织含量最多。缺少该酶容易导致雷-尼二氏综合征,表现为尿酸增高及神经异常,亦称自毁容貌症。但由于目前还没有理想的HPRT基因缺陷动物模型,因此对雷-尼二氏综合征的研究只停留在培养神经细胞水平。

而如今,尹明茹和姜伟华在研究组前期建立的家兔体细胞克隆技术平台基础上,获得了HPRT基因敲除的克隆兔。为了提高基因靶向效率,研究人员使用了基因捕获和rAAV介导的HPRT同源重组设计。本研究为首次在家兔利用同源重组基因打靶技术获得成功,为雷-尼二氏综合征的研究提供了动物模型。本技术也可以应用于家兔其他基因的修饰以及提高其他物种的基因修饰效率。

原文链接:

Generation of hypoxanthine phosphoribosyltransferase gene knockout rabbits by homologous recombination and gene trapping through somatic cell nuclear transfer

原文摘要:

The rabbit is a common animal model that has been employed in studies on various human disorders, and the generation of genetically modified rabbit lines is highly desirable. Female rabbits have been successfully cloned from cumulus cells, and the somatic cell nuclear transfer (SCNT) technology is well established. The present study generated hypoxanthine phosphoribosyltransferase (HPRT) gene knockout rabbits using recombinant adeno-associated virus-mediated homologous recombination and SCNT. Gene trap strategies were employed to enhance the gene targeting rates. The male and female gene knockout fibroblast cell lines were derived by different strategies. When male HPRT knockout cells were used for SCNT, no live rabbits were obtained. However, when female HPRT+/− cells were used for SCNT, live, healthy rabbits were generated. The cloned HPRT+/− rabbits were fertile at maturity. We demonstrate a new technique to produce gene-targeted rabbits. This approach may also be used in the genetic manipulation of different genes or in other species.

来源: Scientific Reports 浏览次数:0

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