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

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...

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Autores principales: Yamamoto, Yasuhiro, Watanabe, Toshiaki, Hoki, Yuko, Shirane, Kenjiro, Li, Yufeng, Ichiiyanagi, Kenji, Kuramochi-Miyagawa, Satomi, Toyoda, Atsushi, Fujiyama, Asao, Oginuma, Masayuki, Suzuki, Hitomi, Sado, Takashi, Nakano, Toru, Sasaki, Hiroyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561870/
https://www.ncbi.nlm.nih.gov/pubmed/23132912
http://dx.doi.org/10.1101/gr.137224.112
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author Yamamoto, Yasuhiro
Watanabe, Toshiaki
Hoki, Yuko
Shirane, Kenjiro
Li, Yufeng
Ichiiyanagi, Kenji
Kuramochi-Miyagawa, Satomi
Toyoda, Atsushi
Fujiyama, Asao
Oginuma, Masayuki
Suzuki, Hitomi
Sado, Takashi
Nakano, Toru
Sasaki, Hiroyuki
author_facet Yamamoto, Yasuhiro
Watanabe, Toshiaki
Hoki, Yuko
Shirane, Kenjiro
Li, Yufeng
Ichiiyanagi, Kenji
Kuramochi-Miyagawa, Satomi
Toyoda, Atsushi
Fujiyama, Asao
Oginuma, Masayuki
Suzuki, Hitomi
Sado, Takashi
Nakano, Toru
Sasaki, Hiroyuki
author_sort Yamamoto, Yasuhiro
collection PubMed
description 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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spelling pubmed-35618702013-08-01 Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus Yamamoto, Yasuhiro Watanabe, Toshiaki Hoki, Yuko Shirane, Kenjiro Li, Yufeng Ichiiyanagi, Kenji Kuramochi-Miyagawa, Satomi Toyoda, Atsushi Fujiyama, Asao Oginuma, Masayuki Suzuki, Hitomi Sado, Takashi Nakano, Toru Sasaki, Hiroyuki Genome Res Research 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. Cold Spring Harbor Laboratory Press 2013-02 /pmc/articles/PMC3561870/ /pubmed/23132912 http://dx.doi.org/10.1101/gr.137224.112 Text en © 2013, Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/3.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported License), as described at http://creativecommons.org/licenses/by-nc/3.0/.
spellingShingle Research
Yamamoto, Yasuhiro
Watanabe, Toshiaki
Hoki, Yuko
Shirane, Kenjiro
Li, Yufeng
Ichiiyanagi, Kenji
Kuramochi-Miyagawa, Satomi
Toyoda, Atsushi
Fujiyama, Asao
Oginuma, Masayuki
Suzuki, Hitomi
Sado, Takashi
Nakano, Toru
Sasaki, Hiroyuki
Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
title Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
title_full Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
title_fullStr Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
title_full_unstemmed Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
title_short Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus
title_sort targeted gene silencing in mouse germ cells by insertion of a homologous dna into a pirna generating locus
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561870/
https://www.ncbi.nlm.nih.gov/pubmed/23132912
http://dx.doi.org/10.1101/gr.137224.112
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