Research

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

    • 1Division of Epigenomics, Department of Molecular Genetics, Medical Institute of Bioregulation, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan;
    • 2Division of Human Genetics, Department of Integrated Genetics, National Institute of Genetics, Research Organization of Information and Systems, Mishima, Shizuoka 411-8540, Japan;
    • 3Department of Cell Biology, School of Medicine, Yale University, New Haven, Connecticut 06520, USA;
    • 4Graduate School of Frontier Biosciences, and Graduate School of Medical Sciences, Osaka University, Suita, Osaka 565-0871, Japan;
    • 5Comparative Genomics Laboratory, Center for Genetic Resource Information, National Institute of Genetics, Research Organization of Information and Systems, Mishima, Shizuoka 411-8540, Japan;
    • 6Mammalian Development Laboratory, Genetic Strains Research Center, National Institute of Genetics, Research Organization of Information and Systems, Mishima, Shizuoka 411-8540, Japan
    • 7 These authors contributed equally to this work.
    • 8 Corresponding author E-mail [email protected]
Published November 6, 2012. Vol 23 Issue 2, pp. 292-299. https://doi.org/10.1101/gr.137224.112
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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.

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