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

Chikara Takeuchi; Kensaku Murano; Mitsuru Ishikawa; Hideyuki Okano; Yuka W. Iwasaki

doi:10.1007/978-1-0716-2380-0_9

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

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

Research output: Chapter in Book/Report/Conference proceeding › Chapter

3 Citations (Scopus)

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 language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	143-153
Number of pages	11
DOIs	https://doi.org/10.1007/978-1-0716-2380-0_9
Publication status	Published - 2022
Externally published	Yes

Publication series

Name	Methods in Molecular Biology
Volume	2509
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

All Science Journal Classification (ASJC) codes

Molecular Biology
Genetics

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10.1007/978-1-0716-2380-0_9

Cite this

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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.",

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doi = "10.1007/978-1-0716-2380-0_9",

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AU - Takeuchi, Chikara

AU - Murano, Kensaku

AU - Ishikawa, Mitsuru

AU - Okano, Hideyuki

AU - Iwasaki, Yuka W.

PY - 2022

Y1 - 2022

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AB - 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.

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

Abstract

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