Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus

Yasuhiro Yamamoto, Toshiaki Watanabe, Yuko Hoki, Kenjiro Shirane, Yufeng Li, Kenji Ichiiyanagi, Satomi Kuramochi-Miyagawa, Atsushi Toyoda, Asao Fujiyama, Masayuki Oginuma, Hitomi Suzuki, Takashi Sado, Toru Nakano, Hiroyuki Sasaki

    Research output: Contribution to journalArticle

    21 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)292-299
    Number of pages8
    JournalGenome Research
    Volume23
    Issue number2
    DOIs
    Publication statusPublished - Feb 2013

    All Science Journal Classification (ASJC) codes

    • Genetics
    • Genetics(clinical)

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  • Cite this

    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. (2013). Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus. Genome Research, 23(2), 292-299. https://doi.org/10.1101/gr.137224.112