Generation of Hprt-disrupted rat through mouse←rat ES chimeras.
Generation of Hprt-disrupted rat through mouse←rat ES chimeras.
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DOI：
10.1038/srep24215
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发表时间：
2016-04-11
影响因子：
4.6
通讯作者：
Ikawa M
中科院分区：
文献类型：
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作者：
Isotani A;Yamagata K;Okabe M;Ikawa M
我们从双转基因大鼠系建立了大鼠胚胎干（ES）细胞系，所述双转基因大鼠系具有用于在体细胞中普遍表达GFP的CAG-GFP和用于在精子中表达Acr 3-EGFP（绿色体和绿色精子：GBGS大鼠）。通过将GBGS大鼠ES细胞注射到小鼠囊胚中并将其移植到假孕小鼠中，在小鼠←大鼠ES嵌合体中产生大鼠精子。通过睾丸精子提取结合卵胞浆内单精子注射（TESE-ICSI），嵌合睾丸的大鼠精子能够使卵子受精。本研究利用小鼠←大鼠ES嵌合体系统，在ES细胞中破坏大鼠次黄嘌呤-鸟嘌呤磷酸核糖转移酶（Hprt）基因，建立了Hprt基因破坏的大鼠系。小鼠←大鼠ES嵌合体系统展示了空间保护和在敲除大鼠生产中明确指示生殖系传递的双重优势。
We established rat embryonic stem (ES) cell lines from a double transgenic rat line which harbours CAG-GFP for ubiquitous expression of GFP in somatic cells and Acr3-EGFP for expression in sperm (green body and green sperm: GBGS rat). By injecting the GBGS rat ES cells into mouse blastocysts and transplanting them into pseudopregnant mice, rat spermatozoa were produced in mouse←rat ES chimeras. Rat spermatozoa from the chimeric testis were able to fertilize eggs by testicular sperm extraction combined with intracytoplasmic sperm injection (TESE-ICSI). In the present paper, we disrupted rat hypoxanthine-guanine phosphoribosyl transferase (Hprt) gene in ES cells and produced a Hprt-disrupted rat line using the mouse←rat ES chimera system. The mouse←rat ES chimera system demonstrated the dual advantages of space conservation and a clear indication of germ line transmission in knockout rat production.