datacite.contributor.supervisor	editor: Bronner, Marianne E
datacite.contributor.supervisor	senior_editor: Bronner, Marianne E
dc.contributor.author	Sun, Dawei
dc.contributor.author	Evans, Lewis
dc.contributor.author	Perrone, Francesca
dc.contributor.author	Sokleva, Vanesa
dc.contributor.author	Lim, Kyungtae
dc.contributor.author	Rezakhani, Saba
dc.contributor.author	Lutolf, Matthias
dc.contributor.author	Zilbauer, Matthias
dc.contributor.author	Rawlins, Emma L
dc.contributor.orcid	Sun, Dawei [0000-0003-1551-4349]
dc.contributor.orcid	Evans, Lewis [0000-0001-7279-7651]
dc.contributor.orcid	Lim, Kyungtae [0000-0001-6044-2191]
dc.contributor.orcid	Rawlins, Emma L [0000-0001-7426-3792]
dc.date.accessioned	2021-10-29T02:13:16Z
dc.date.available	2021-10-29T02:13:16Z
dc.date.issued	2021-10-06
dc.date.submitted	2021-03-03
dc.date.updated	2021-10-29T02:13:13Z
dc.description	Funder: Alzheimers Research UK Stem Cell Research Centre
dc.description.abstract	Human organoid systems recapitulate key features of organs offering platforms for modelling developmental biology and disease. Tissue-derived organoids have been widely used to study the impact of extrinsic niche factors on stem cells. However, they are rarely used to study endogenous gene function due to the lack of efficient gene manipulation tools. Previously, we established a human foetal lung organoid system (Nikolić et al., 2017). Here, using this organoid system as an example, we have systematically developed and optimised a complete genetic toolbox for use in tissue-derived organoids. This includes ‘Organoid Easytag’, our efficient workflow for targeting all types of gene loci through CRISPR-mediated homologous recombination followed by flow cytometry for enriching correctly targeted cells. Our toolbox also incorporates conditional gene knockdown or overexpression using tightly inducible CRISPR interference and CRISPR activation which is the first efficient application of these techniques to tissue-derived organoids. These tools will facilitate gene perturbation studies in tissue-derived organoids facilitating human disease modelling and providing a functional counterpart to many ongoing descriptive studies, such as the Human Cell Atlas Project.
dc.identifier.doi	10.17863/CAM.77469
dc.identifier.eissn	2050-084X
dc.identifier.other	67886
dc.identifier.uri	https://www.repository.cam.ac.uk/handle/1810/330025
dc.language	en
dc.publisher	eLife Sciences Publications, Ltd
dc.subject	Research Advance
dc.subject	Developmental Biology
dc.subject	organoids
dc.subject	lung
dc.subject	CRISPR-Cas9
dc.subject	homologous gene targeting
dc.subject	inducible CRISPRi
dc.subject	CRISPRa
dc.subject	Human
dc.title	A functional genetic toolbox for human tissue-derived organoids
dc.type	Article
dcterms.dateAccepted	2021-10-05
prism.publicationName	eLife
prism.volume	10
pubs.funder-project-id	Medical Research Council (MR/P009581/1)
pubs.funder-project-id	Wellcome Trust (PhD Studentship 109146/Z/15/Z)
pubs.funder-project-id	National Research Foundation of Korea (2018R1A6A3A03012122)
pubs.funder-project-id	Wellcome Trust (Core Support for Gurdon Institute 203144/Z/16/Z)
pubs.funder-project-id	Cancer Research UK (Core Support for Gurdon Institute C6946/A24843)
pubs.funder-project-id	Medical Research Council (New Investigator Research Grant MR/T001917/1)
pubs.funder-project-id	Wellcome Trust (PhD studentship 102175/B/13/Z)
rioxxterms.licenseref.uri	http://creativecommons.org/licenses/by/4.0/
rioxxterms.version	VoR
rioxxterms.versionofrecord	10.7554/elife.67886

A functional genetic toolbox for human tissue-derived organoids

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