Imperial College London
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Dr Kevin Woollard

Faculty of Medicine, Department of Immunology and Inflammation

Honorary Senior Lecturer
 
 
 
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Contact

 

+44 (0)20 3313 2357k.woollard Website

 
 
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Location

 

9N15CCommonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Shamshirgaran:2021:10.1038/s41598-021-95986-5,
author = {Shamshirgaran, Y and Jonebring, A and Svensson, A and Leefa, I and Bohlooly-Y, M and Firth, M and Woollard, KJ and Hofherr, A and Rogers, IM and Hicks, R},
doi = {10.1038/s41598-021-95986-5},
journal = {Scientific Reports},
pages = {1--9},
title = {Rapid target validation in a Cas9-inducible hiPSC derived kidney model},
url = {http://dx.doi.org/10.1038/s41598-021-95986-5},
volume = {11},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - 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.
AU  - Shamshirgaran,Y
AU  - Jonebring,A
AU  - Svensson,A
AU  - Leefa,I
AU  - Bohlooly-Y,M
AU  - Firth,M
AU  - Woollard,KJ
AU  - Hofherr,A
AU  - Rogers,IM
AU  - Hicks,R
DO  - 10.1038/s41598-021-95986-5
EP  - 9
PY  - 2021///
SN  - 2045-2322
SP  - 1
TI  - Rapid target validation in a Cas9-inducible hiPSC derived kidney model
T2  - Scientific Reports
UR  - http://dx.doi.org/10.1038/s41598-021-95986-5
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000686663200047&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - https://www.nature.com/articles/s41598-021-95986-5
UR  - http://hdl.handle.net/10044/1/92416
VL  - 11
ER  -
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