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通过卵母细胞注射使用转录激活样效应因子核酸酶(TALEN)制备Sry基因敲除小鼠。

Production of Sry knockout mouse using TALEN via oocyte injection.

作者信息

Kato Tomoko, Miyata Kohei, Sonobe Miku, Yamashita Satoshi, Tamano Moe, Miura Kento, Kanai Yoshiakira, Miyamoto Shingo, Sakuma Tetsushi, Yamamoto Takashi, Inui Masafumi, Kikusui Takefumi, Asahara Hiroshi, Takada Shuji

机构信息

1] Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan [2].

出版信息

Sci Rep. 2013 Nov 5;3:3136. doi: 10.1038/srep03136.

Abstract

Recently developed transcription activator-like effector nuclease (TALEN) technology has enabled the creation of knockout mice, even for genes on the Y chromosome. In this study, we generated a knockout mouse for Sry, a sex-determining gene on the Y chromosome, using microinjection of TALEN RNA into pronuclear stage oocytes. As expected, the knockout mouse had female external and internal genitalia, a female level of blood testosterone and a female sexually dimorphic nucleus in the brain. The knockout mouse exhibited an estrous cycle and performed copulatory behavior as females, although it was infertile or had reduced fertility. A histological analysis showed that the ovary of the knockout mouse displayed a reduced number of oocytes and luteinized unruptured follicles, implying that a reduced number of ovulated oocytes is a possible reason for infertility and/or reduced fertility in the KO mouse.

摘要

最近开发的转录激活样效应核酸酶(TALEN)技术使得敲除小鼠的基因成为可能,即使是Y染色体上的基因。在本研究中,我们通过向原核期卵母细胞显微注射TALEN RNA,构建了Y染色体上的性别决定基因Sry的敲除小鼠。正如预期的那样,该敲除小鼠具有雌性的外部和内部生殖器、雌性水平的血液睾酮以及大脑中的雌性性二态核。该敲除小鼠表现出发情周期,并表现出雌性的交配行为,尽管它不育或生育力降低。组织学分析表明,敲除小鼠的卵巢中卵母细胞数量减少,出现了黄素化未破裂卵泡,这意味着排卵卵母细胞数量减少可能是敲除小鼠不育和/或生育力降低的一个原因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9fc5/3817445/e791b7a0bf95/srep03136-f1.jpg

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