Identification of mouse piRNA pathway components using anti-MIWI2 antibodies

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2 Citations (Scopus)

Abstract

The mouse testis has served as a popular model system to study a wide range of biological processes, including germcell development, meiosis, epigenetic changes of chromatin, transposon silencing, and small RNA-mediated epigenetic modifications. PIWI-interacting RNAs (piRNAs) are a class of small RNAs that are almost exclusively expressed in animal gonads. They repress transposons by forming effector complexes with PIWI proteins to maintain genome integrity of the germline. Here we describe detailed procedures of how to produce monoclonal antibodies against a mouse nuclear PIWI protein, MIWI2, which functions in de novo DNA methylation of target transposon loci. We then describe how to use the antibodies to isolate associated complexes and to detect MIWI2 immunohistochemically.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages205-216
Number of pages12
Volume1463
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1463
ISSN (Print)10643745

Keywords

  • Inclusion body
  • MIWI2
  • Monoclonal antibody
  • Mouse embryonic testes
  • piRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Hirano, T., Hasuwa, H., & Siomi, H. (2017). Identification of mouse piRNA pathway components using anti-MIWI2 antibodies. In Methods in Molecular Biology (Vol. 1463, pp. 205-216). (Methods in Molecular Biology; Vol. 1463). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-4017-2_15