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

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

. 2022:2509:143-153.

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

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

Chikara Takeuchi¹, Kensaku Murano¹, Mitsuru Ishikawa², Hideyuki Okano², Yuka W Iwasaki^{3

4}

Affiliations

¹ Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
² Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
³ Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan. iwasaki@keio.jp.
⁴ Japan Science and Technology Agency (JST), Precursory Research for Embryonic Science and Technology (PRESTO), Saitama, Japan. iwasaki@keio.jp.

PMID: 35796962
DOI: 10.1007/978-1-0716-2380-0_9

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

Chikara Takeuchi et al. Methods Mol Biol. 2022.

. 2022:2509:143-153.

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

Authors

Chikara Takeuchi¹, Kensaku Murano¹, Mitsuru Ishikawa², Hideyuki Okano², Yuka W Iwasaki^{3

4}

Affiliations

¹ Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
² Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
³ Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan. iwasaki@keio.jp.
⁴ Japan Science and Technology Agency (JST), Precursory Research for Embryonic Science and Technology (PRESTO), Saitama, Japan. iwasaki@keio.jp.

PMID: 35796962
DOI: 10.1007/978-1-0716-2380-0_9

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.

Keywords: Ovarian somatic cell; PIWI; Stable line; piRNA pathway; piggyBac.

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References

1. Ozata DM, Gainetdinov I, Zoch A, O'Carroll D, Zamore PD (2019) PIWI-interacting RNAs: small RNAs with big functions. Nat Rev Genet 20(2):89–108. https://doi.org/10.1038/s41576-018-0073-3 - DOI - PubMed
1. Iwasaki YW, Siomi MC, Siomi H (2015) PIWI-interacting RNA: its biogenesis and functions. Annu Rev Biochem 84:405–433. https://doi.org/10.1146/annurev-biochem-060614-034258 - DOI - PubMed
1. Senti KA, Brennecke J (2010) The piRNA pathway: a fly's perspective on the guardian of the genome. Trends Genet 26(12):499–509. https://doi.org/10.1016/j.tig.2010.08.007 - DOI - PubMed - PMC
1. Niki Y, Yamaguchi T, Mahowald AP (2006) Establishment of stable cell lines of Drosophila germ-line stem cells. Proc Natl Acad Sci U S A 103(44):16325–16330. https://doi.org/10.1073/pnas.0607435103 - DOI - PubMed - PMC
1. Saito K, Inagaki S, Mituyama T, Kawamura Y, Ono Y, Sakota E, Kotani H, Asai K, Siomi H, Siomi MC (2009) A regulatory circuit for piwi by the large Maf gene traffic jam in drosophila. Nature 461(7268):1296–1299. https://doi.org/10.1038/nature08501 - DOI - PubMed

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[1] Ozata DM, Gainetdinov I, Zoch A, O'Carroll D, Zamore PD (2019) PIWI-interacting RNAs: small RNAs with big functions. Nat Rev Genet 20(2):89–108. https://doi.org/10.1038/s41576-018-0073-3 - DOI - PubMed

[2] Ozata DM, Gainetdinov I, Zoch A, O'Carroll D, Zamore PD (2019) PIWI-interacting RNAs: small RNAs with big functions. Nat Rev Genet 20(2):89–108. https://doi.org/10.1038/s41576-018-0073-3 - DOI - PubMed

[3] Iwasaki YW, Siomi MC, Siomi H (2015) PIWI-interacting RNA: its biogenesis and functions. Annu Rev Biochem 84:405–433. https://doi.org/10.1146/annurev-biochem-060614-034258 - DOI - PubMed

[4] Iwasaki YW, Siomi MC, Siomi H (2015) PIWI-interacting RNA: its biogenesis and functions. Annu Rev Biochem 84:405–433. https://doi.org/10.1146/annurev-biochem-060614-034258 - DOI - PubMed

[5] Senti KA, Brennecke J (2010) The piRNA pathway: a fly's perspective on the guardian of the genome. Trends Genet 26(12):499–509. https://doi.org/10.1016/j.tig.2010.08.007 - DOI - PubMed - PMC

[6] Senti KA, Brennecke J (2010) The piRNA pathway: a fly's perspective on the guardian of the genome. Trends Genet 26(12):499–509. https://doi.org/10.1016/j.tig.2010.08.007 - DOI - PubMed - PMC

[7] Niki Y, Yamaguchi T, Mahowald AP (2006) Establishment of stable cell lines of Drosophila germ-line stem cells. Proc Natl Acad Sci U S A 103(44):16325–16330. https://doi.org/10.1073/pnas.0607435103 - DOI - PubMed - PMC

[8] Niki Y, Yamaguchi T, Mahowald AP (2006) Establishment of stable cell lines of Drosophila germ-line stem cells. Proc Natl Acad Sci U S A 103(44):16325–16330. https://doi.org/10.1073/pnas.0607435103 - DOI - PubMed - PMC

[9] Saito K, Inagaki S, Mituyama T, Kawamura Y, Ono Y, Sakota E, Kotani H, Asai K, Siomi H, Siomi MC (2009) A regulatory circuit for piwi by the large Maf gene traffic jam in drosophila. Nature 461(7268):1296–1299. https://doi.org/10.1038/nature08501 - DOI - PubMed

[10] Saito K, Inagaki S, Mituyama T, Kawamura Y, Ono Y, Sakota E, Kotani H, Asai K, Siomi H, Siomi MC (2009) A regulatory circuit for piwi by the large Maf gene traffic jam in drosophila. Nature 461(7268):1296–1299. https://doi.org/10.1038/nature08501 - DOI - PubMed

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

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

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