A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B

Abstract Genome-wide RNA interference (RNAi) with pooled and barcoded short-hairpin RNA (shRNA) libraries provides a powerful tool for identifying cellular components that are relevant to the modes/mechanisms of action (MoA) of bioactive compounds. shRNAs that affect cellular sensitivity to a given...

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Autores principales:	Shohei Takase, Rumi Kurokawa, Daisuke Arai, Kind Kanemoto Kanto, Tatsufumi Okino, Yoichi Nakao, Tetsuo Kushiro, Minoru Yoshida, Ken Matsumoto
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ebfbac593a8b4e36ad2dc6b7dd41f0e4
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spelling	oai:doaj.org-article:ebfbac593a8b4e36ad2dc6b7dd41f0e42021-12-02T12:30:27ZA quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B10.1038/s41598-017-02016-42045-2322https://doaj.org/article/ebfbac593a8b4e36ad2dc6b7dd41f0e42017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02016-4https://doaj.org/toc/2045-2322Abstract Genome-wide RNA interference (RNAi) with pooled and barcoded short-hairpin RNA (shRNA) libraries provides a powerful tool for identifying cellular components that are relevant to the modes/mechanisms of action (MoA) of bioactive compounds. shRNAs that affect cellular sensitivity to a given compound can be identified by deep sequencing of shRNA-specific barcodes. We used multiplex barcode sequencing technology by adding sample-specific index tags to PCR primers during sequence library preparation, enabling parallel analysis of multiple samples. An shRNA library screen with this system revealed that downregulation of ATP1A1, an α-subunit of Na+/K+ ATPase, conferred significant sensitivity to aurilide B, a natural marine product that induces mitochondria-mediated apoptosis. Combined treatment with ouabain which inhibits Na+/K+ ATPase by targeting α-subunits potentiated sensitivity to aurilide B, suggesting that ATP1A1 regulates mitochondria-mediated apoptosis. Our results indicate that multiplex sequencing facilitates the use of pooled shRNA library screening for the identification of combination drug therapy targets.Shohei TakaseRumi KurokawaDaisuke AraiKind Kanemoto KantoTatsufumi OkinoYoichi NakaoTetsuo KushiroMinoru YoshidaKen MatsumotoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shohei Takase Rumi Kurokawa Daisuke Arai Kind Kanemoto Kanto Tatsufumi Okino Yoichi Nakao Tetsuo Kushiro Minoru Yoshida Ken Matsumoto A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B
description	Abstract Genome-wide RNA interference (RNAi) with pooled and barcoded short-hairpin RNA (shRNA) libraries provides a powerful tool for identifying cellular components that are relevant to the modes/mechanisms of action (MoA) of bioactive compounds. shRNAs that affect cellular sensitivity to a given compound can be identified by deep sequencing of shRNA-specific barcodes. We used multiplex barcode sequencing technology by adding sample-specific index tags to PCR primers during sequence library preparation, enabling parallel analysis of multiple samples. An shRNA library screen with this system revealed that downregulation of ATP1A1, an α-subunit of Na+/K+ ATPase, conferred significant sensitivity to aurilide B, a natural marine product that induces mitochondria-mediated apoptosis. Combined treatment with ouabain which inhibits Na+/K+ ATPase by targeting α-subunits potentiated sensitivity to aurilide B, suggesting that ATP1A1 regulates mitochondria-mediated apoptosis. Our results indicate that multiplex sequencing facilitates the use of pooled shRNA library screening for the identification of combination drug therapy targets.
format	article
author	Shohei Takase Rumi Kurokawa Daisuke Arai Kind Kanemoto Kanto Tatsufumi Okino Yoichi Nakao Tetsuo Kushiro Minoru Yoshida Ken Matsumoto
author_facet	Shohei Takase Rumi Kurokawa Daisuke Arai Kind Kanemoto Kanto Tatsufumi Okino Yoichi Nakao Tetsuo Kushiro Minoru Yoshida Ken Matsumoto
author_sort	Shohei Takase
title	A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B
title_short	A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B
title_full	A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B
title_fullStr	A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B
title_full_unstemmed	A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B
title_sort	quantitative shrna screen identifies atp1a1 as a gene that regulates cytotoxicity by aurilide b
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/ebfbac593a8b4e36ad2dc6b7dd41f0e4
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A quantitative shRNA screen identifies ATP1A1 as a gene that regulates cytotoxicity by aurilide B

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