Genetic Modification and Screening in Rat Using Haploid Embryonic Stem Cells
Genetic Modification and Screening in Rat Using Haploid Embryonic Stem Cells
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DOI：
10.1016/j.stem.2013.11.016
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发表时间：
2014-03-06
期刊：
影响因子：
23.9
通讯作者：
Zhou, Qi
中科院分区：
文献类型：
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作者：
Li, Wei;Li, Xin;Zhou, Qi
大鼠是生物医学研究中重要的动物模型，但遗传操作的实际局限性限制了遗传分析的应用。在这里，我们报告的衍生大鼠雄激素单倍体胚胎干细胞（RahESCs）作为一种工具，以促进此类研究。我们的方法是基于从受精卵中去除母体原核以产生雄核发育胚胎，然后衍生ESC。所得到的RahESC具有21条染色体，表达多能性标记，分化成三个胚层细胞，并有助于种系。纯合突变可以通过同源重组或CRISPR-Cas系统通过大规模基因捕获和精确基因靶向引入。RahESC也可以在卵母细胞胞质内注射后产生可生育的大鼠，因此能够将遗传修饰传递给后代。总体而言，RahESC代表了大鼠功能遗传学研究和转基因品系生产的实用工具。
The rat is an important animal model in biomedical research, but practical limitations to genetic manipulation have restricted the application of genetic analysis. Here we report the derivation of rat androgenetic haploid embryonic stem cells (RahESCs) as a tool to facilitate such studies. Our approach is based on removal of the maternal pronucleus from zygotes to generate androgenetic embryos followed by derivation of ESCs. The resulting RahESCs have 21 chromosomes, express pluripotency markers, differentiate into three germ layer cells, and contribute to the germline. Homozygous mutations can be introduced by both large-scale gene trapping and precise gene targeting via homologous recombination or the CRISPR-Cas system. RahESCs can also produce fertile rats after intracytoplasmic injection into oocytes and are therefore able to transmit genetic modifications to offspring. Overall, RahESCs represent a practical tool for functional genetic studies and production of transgenic lines in rat.