Identification of Mouse piRNA Pathway Components Using Anti-MIWI2 Antibodies
The mouse testis has served as a popular model system to study a wide range of biological processes, including germ cell 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.
Key wordspiRNA MIWI2 Monoclonal antibody Inclusion body Mouse embryonic testes
The authors would like to thank the members of the Siomi laboratory for their comments and for critical reading of the manuscript. This work was supported by grants from MEXT to H.S.
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