Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus.
Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus.
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DOI：
10.1101/gr.137224.112
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发表时间：
2013-02
期刊：
影响因子：
7
通讯作者：
Sasaki H
中科院分区：
文献类型：
--
作者：
Yamamoto Y;Watanabe T;Hoki Y;Shirane K;Li Y;Ichiiyanagi K;Kuramochi-Miyagawa S;Toyoda A;Fujiyama A;Oginuma M;Suzuki H;Sado T;Nakano T;Sasaki H
在动物的生殖细胞、早期胚胎和干细胞中，PIWI 相互作用 RNA (piRNA) 通过转录和转录后机制，在沉默逆转录转座子（一种恶性基因组寄生虫）方面发挥着重要作用。为了检查 piRNA 途径是否可用于沉默生殖细胞中感兴趣的基因，我们培育了基因敲入小鼠，其中将外源 DNA 片段插入产生粗线期 piRNA 的区域。敲入序列被转录，产生的 RNA 被加工以在出生后睾丸中产生 piRNA。当引入具有与部分敲入序列互补的序列的报告基因时，它们在粗线期piRNA生成后被极大地抑制。这种抑制主要发生在转录后水平，因为报告 RNA 的降解加速。我们的结果表明，piRNA 途径可用作生殖细胞中序列特异性基因沉默的工具，并支持 piRNA 生成区域作为逆转录转座子陷阱的观点，使宿主细胞能够生成针对活性逆转录转座子的 piRNA。
In germ cells, early embryos, and stem cells of animals, PIWI-interacting RNAs (piRNAs) have an important role in silencing retrotransposons, which are vicious genomic parasites, through transcriptional and post-transcriptional mechanisms. To examine whether the piRNA pathway can be used to silence genes of interest in germ cells, we have generated knock-in mice in which a foreign DNA fragment was inserted into a region generating pachytene piRNAs. The knock-in sequence was transcribed, and the resulting RNA was processed to yield piRNAs in postnatal testes. When reporter genes possessing a sequence complementary to portions of the knock-in sequence were introduced, they were greatly repressed after the time of pachytene piRNA generation. This repression mainly occurred at the post-transcriptional level, as degradation of the reporter RNAs was accelerated. Our results show that the piRNA pathway can be used as a tool for sequence-specific gene silencing in germ cells and support the idea that the piRNA generating regions serve as traps for retrotransposons, enabling the host cell to generate piRNAs against active retrotransposons.
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影响因子：
4.1
作者：
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通讯作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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