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

Academic Article

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. © 2013, Published by Cold Spring Harbor Laboratory Press.
  • Authors

    Digital Object Identifier (doi)

    Author List

  • Yamamoto Y; Watanabe T; Hoki Y; Shirane K; Li Y; Ichiiyanagi K; Kuramochi-Miyagawa S; Toyoda A; Fujiyama A; Oginuma M
  • Start Page

  • 292
  • End Page

  • 299
  • Volume

  • 23
  • Issue

  • 2