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Organoid Easytag: an efficient workflow for gene targeting in human organoids

View ORCID ProfileDawei Sun, View ORCID ProfileLewis D. Evans, View ORCID ProfileEmma L. Rawlins
doi: https://doi.org/10.1101/2020.05.04.076067
Dawei Sun
1Wellcome Trust/CRUK Gurdon Institute, University of Cambridge, Cambridge, CB2 1QN, UK
2Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, UK
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Lewis D. Evans
3Developmental Biology and Cancer Department, University College London, London, WC1N 1DZ, UK
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Emma L. Rawlins
1Wellcome Trust/CRUK Gurdon Institute, University of Cambridge, Cambridge, CB2 1QN, UK
4Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EL, UK
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  • For correspondence: e.rawlins@gurdon.cam.ac.uk
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Abstract

Human organoid systems recapitulate key features of organs offering platforms for modelling human developmental biology and disease. However, tissue-derived organoids suffer from low efficiency of genetic manipulations, especially CRISPR/Cas9-mediated knock-ins. We have systematically optimised and developed an “Organoid Easytag” pipeline for efficient (40-65%) and accurate gene targeting to facilitate generation of reporter lines and gene knock-outs in organoid based research.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Contact: e.rawlins{at}gurdon.cam.ac.uk

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 05, 2020.
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Organoid Easytag: an efficient workflow for gene targeting in human organoids
Dawei Sun, Lewis D. Evans, Emma L. Rawlins
bioRxiv 2020.05.04.076067; doi: https://doi.org/10.1101/2020.05.04.076067
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Organoid Easytag: an efficient workflow for gene targeting in human organoids
Dawei Sun, Lewis D. Evans, Emma L. Rawlins
bioRxiv 2020.05.04.076067; doi: https://doi.org/10.1101/2020.05.04.076067

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