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dc.contributor.authorSun, Dawei
dc.contributor.authorEvans, Lewis
dc.contributor.authorPerrone, Francesca
dc.contributor.authorSokleva, Vanesa
dc.contributor.authorLim, Kyungtae
dc.contributor.authorRezakhani, Saba
dc.contributor.authorLutolf, Matthias
dc.contributor.authorZilbauer, Matthias
dc.contributor.authorRawlins, Emma
dc.date.accessioned2021-10-07T23:30:57Z
dc.date.available2021-10-07T23:30:57Z
dc.date.issued2021-10-06
dc.identifier.issn2050-084X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/329127
dc.description.abstractHuman 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.format.mediumPrint-Electronic
dc.languageeng
dc.publishereLife Sciences Publications, Ltd
dc.rightsAll rights reserved
dc.titleA functional genetic toolbox for human tissue-derived organoids.
dc.typeArticle
prism.publicationDate2021
prism.publicationNameElife
prism.volume10
dc.identifier.doi10.17863/CAM.76573
dcterms.dateAccepted2021-10-05
rioxxterms.versionofrecord10.7554/eLife.67886
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2021-10-06
dc.contributor.orcidSun, Dawei [0000-0003-1551-4349]
dc.contributor.orcidEvans, Lewis [0000-0001-7279-7651]
dc.contributor.orcidLim, Kyungtae [0000-0001-6044-2191]
dc.contributor.orcidZilbauer, Matthias [0000-0002-7272-0547]
dc.contributor.orcidRawlins, Emma [0000-0001-7426-3792]
dc.identifier.eissn2050-084X
rioxxterms.typeJournal Article/Review
pubs.funder-project-idMedical Research Council (MR/P009581/1)
pubs.funder-project-idMRC (MR/T001917/1)
pubs.funder-project-idWellcome Trust (109146/Z/15/Z)
pubs.funder-project-idCancer Research UK (C6946/A24843)
pubs.funder-project-idWellcome Trust (203144/Z/16/Z)
pubs.funder-project-idWellcome Trust (102175/B/13/Z)
cam.issuedOnline2021-10-06
rioxxterms.freetoread.startdate2024-10-07


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