Rapid target validation in a Cas9-inducible hiPSC derived kidney model

Abstract Recent advances in induced pluripotent stem cells (iPSCs), genome editing technologies and 3D organoid model systems highlight opportunities to develop new in vitro human disease models to serve drug discovery programs. An ideal disease model would accurately recapitulate the relevant disea...

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Autores principales:	Yasaman Shamshirgaran, Anna Jonebring, Anna Svensson, Isabelle Leefa, Mohammad Bohlooly-Y, Mike Firth, Kevin J. Woollard, Alexis Hofherr, Ian M. Rogers, Ryan Hicks
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/671fd9f7c0264f90af36569f226a7d4c
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spelling	oai:doaj.org-article:671fd9f7c0264f90af36569f226a7d4c2021-12-02T16:45:46ZRapid target validation in a Cas9-inducible hiPSC derived kidney model10.1038/s41598-021-95986-52045-2322https://doaj.org/article/671fd9f7c0264f90af36569f226a7d4c2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95986-5https://doaj.org/toc/2045-2322Abstract Recent advances in induced pluripotent stem cells (iPSCs), genome editing technologies and 3D organoid model systems highlight opportunities to develop new in vitro human disease models to serve drug discovery programs. An ideal disease model would accurately recapitulate the relevant disease phenotype and provide a scalable platform for drug and genetic screening studies. Kidney organoids offer a high cellular complexity that may provide greater insights than conventional single-cell type cell culture models. However, genetic manipulation of the kidney organoids requires prior generation of genetically modified clonal lines, which is a time and labor consuming procedure. Here, we present a methodology for direct differentiation of the CRISPR-targeted cell pools, using a doxycycline-inducible Cas9 expressing hiPSC line for high efficiency editing to eliminate the laborious clonal line generation steps. We demonstrate the versatile use of genetically engineered kidney organoids by targeting the autosomal dominant polycystic kidney disease (ADPKD) genes: PKD1 and PKD2. Direct differentiation of the respective knockout pool populations into kidney organoids resulted in the formation of cyst-like structures in the tubular compartment. Our findings demonstrated that we can achieve > 80% editing efficiency in the iPSC pool population which resulted in a reliable 3D organoid model of ADPKD. The described methodology may provide a platform for rapid target validation in the context of disease modeling.Yasaman ShamshirgaranAnna JonebringAnna SvenssonIsabelle LeefaMohammad Bohlooly-YMike FirthKevin J. WoollardAlexis HofherrIan M. RogersRyan HicksNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yasaman Shamshirgaran Anna Jonebring Anna Svensson Isabelle Leefa Mohammad Bohlooly-Y Mike Firth Kevin J. Woollard Alexis Hofherr Ian M. Rogers Ryan Hicks Rapid target validation in a Cas9-inducible hiPSC derived kidney model
description	Abstract Recent advances in induced pluripotent stem cells (iPSCs), genome editing technologies and 3D organoid model systems highlight opportunities to develop new in vitro human disease models to serve drug discovery programs. An ideal disease model would accurately recapitulate the relevant disease phenotype and provide a scalable platform for drug and genetic screening studies. Kidney organoids offer a high cellular complexity that may provide greater insights than conventional single-cell type cell culture models. However, genetic manipulation of the kidney organoids requires prior generation of genetically modified clonal lines, which is a time and labor consuming procedure. Here, we present a methodology for direct differentiation of the CRISPR-targeted cell pools, using a doxycycline-inducible Cas9 expressing hiPSC line for high efficiency editing to eliminate the laborious clonal line generation steps. We demonstrate the versatile use of genetically engineered kidney organoids by targeting the autosomal dominant polycystic kidney disease (ADPKD) genes: PKD1 and PKD2. Direct differentiation of the respective knockout pool populations into kidney organoids resulted in the formation of cyst-like structures in the tubular compartment. Our findings demonstrated that we can achieve > 80% editing efficiency in the iPSC pool population which resulted in a reliable 3D organoid model of ADPKD. The described methodology may provide a platform for rapid target validation in the context of disease modeling.
format	article
author	Yasaman Shamshirgaran Anna Jonebring Anna Svensson Isabelle Leefa Mohammad Bohlooly-Y Mike Firth Kevin J. Woollard Alexis Hofherr Ian M. Rogers Ryan Hicks
author_facet	Yasaman Shamshirgaran Anna Jonebring Anna Svensson Isabelle Leefa Mohammad Bohlooly-Y Mike Firth Kevin J. Woollard Alexis Hofherr Ian M. Rogers Ryan Hicks
author_sort	Yasaman Shamshirgaran
title	Rapid target validation in a Cas9-inducible hiPSC derived kidney model
title_short	Rapid target validation in a Cas9-inducible hiPSC derived kidney model
title_full	Rapid target validation in a Cas9-inducible hiPSC derived kidney model
title_fullStr	Rapid target validation in a Cas9-inducible hiPSC derived kidney model
title_full_unstemmed	Rapid target validation in a Cas9-inducible hiPSC derived kidney model
title_sort	rapid target validation in a cas9-inducible hipsc derived kidney model
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/671fd9f7c0264f90af36569f226a7d4c
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Rapid target validation in a Cas9-inducible hiPSC derived kidney model

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