Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
- Yasuhiro Yamamoto1,
- Toshiaki Watanabe2,
- Yuko Hoki1,
- Kenjirou Shirane1,
- Yufeng Li1,
- Kenji Ichiiyanagi1,
- Satomi Kuramochi-Miyagawa3,
- Atsushi Toyoda4,
- Asao Fujiyama4,
- Masayuki Oginuma4,
- Hitomi Suzuki4,
- Takashi Sado1,
- Toru Nakano3 and
- Hiroyuki Sasaki1,5
- 1 Division of Epigenomics, Department of Molecular Genetics, Medical Institute of Bioregulation, Kyush;
- 2 Department of Cell Biology, School of Medicine, Yale University;
- 3 raduate School of Frontier Biosciences and Graduate School of Medical Sciences, Osaka University;
- 4 National Institute of Genetics, Research Organization of Information and Systems
- ↵* Corresponding author; email: hsasaki{at}bioreg.kyushu-u.ac.jp
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 posttranscriptional mechanisms. To examine whether the piRNA pathway can be used to silence genes of interest in germ cells, we have generated knockin mice in which a foreign DNA fragment was inserted into a region generating pachytene piRNAs. The knockin 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 knockin sequence were introduced, they were greatly repressed after the time of pachytene piRNA generation. This repression mainly occurred at the posttranscriptional 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.
- Received January 4, 2012.
- Accepted October 23, 2012.
- © 2012, Published by Cold Spring Harbor Laboratory Press
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