Generation of Stable Drosophila Ovarian Somatic Cell Lines Using the piggyBac System

Chikara Takeuchi, Kensaku Murano, Mitsuru Ishikawa, Hideyuki Okano, Yuka Iwasaki

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Transposable elements (TEs) constitute a large proportion of the genome in multiple organisms. Therefore, anti-transposable element machineries are essential to maintain genomic integrity. PIWI-interacting RNAs (piRNAs) are a major force to repress TEs in Drosophila ovaries. Ovarian somatic cells (OSC), in which nuclear piRNA regulation is functional, have been used for research on piRNA pathway as a cell culture system to elucidate the molecular mechanisms underlying the piRNA pathway. Analysis of piRNA pathway using a reporter system to monitor the gene regulation or overexpression of specific genes would be a powerful approach. Here, we present the technical protocol to establish stable cell lines using the piggyBac system, adopted for OSCs. This easy, consistent, and timesaving protocol may accelerate research on the piRNA pathway.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages143-153
Number of pages11
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2509
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Ovarian somatic cell
  • piggyBac
  • piRNA pathway
  • PIWI
  • Stable line

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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