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Rapid, one-step generation of biallelic conditional gene knockouts

Amanda Andersson-Rolf, Roxana C. Mustata, Alessandra Merenda, Sajith Perera, Tiago Grego, Jihoon Kim, Katie Andrews, Juergen Fink, William C. Skarnes, Bon-Kyoung Koo
doi: https://doi.org/10.1101/056549
Amanda Andersson-Rolf
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
2Department of Genetics, University of Cambridge, Cambridge, CB2 3EH, UK.
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Roxana C. Mustata
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
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Alessandra Merenda
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
2Department of Genetics, University of Cambridge, Cambridge, CB2 3EH, UK.
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Sajith Perera
3Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK
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Tiago Grego
3Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK
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Jihoon Kim
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
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Katie Andrews
3Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK
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Juergen Fink
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
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William C. Skarnes
3Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK
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  • For correspondence: bkk25@cam.ac.uk skarnes@sanger.ac.uk
Bon-Kyoung Koo
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
2Department of Genetics, University of Cambridge, Cambridge, CB2 3EH, UK.
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  • For correspondence: bkk25@cam.ac.uk skarnes@sanger.ac.uk
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Abstract

Loss-of-function studies are key to investigate gene function and CRISPR technology has made genome editing widely accessible in model organisms and cells. However, conditional gene inactivation in diploid cells is still difficult to achieve. Here, we present CRISPR-FLIP, a strategy that provides an efficient, rapid, and scalable method for bi-allelic conditional gene knockouts in diploid cells by co-delivery of CRISPR/Cas9 and a universal conditional intron cassette.

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Posted June 01, 2016.
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Rapid, one-step generation of biallelic conditional gene knockouts
Amanda Andersson-Rolf, Roxana C. Mustata, Alessandra Merenda, Sajith Perera, Tiago Grego, Jihoon Kim, Katie Andrews, Juergen Fink, William C. Skarnes, Bon-Kyoung Koo
bioRxiv 056549; doi: https://doi.org/10.1101/056549
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Rapid, one-step generation of biallelic conditional gene knockouts
Amanda Andersson-Rolf, Roxana C. Mustata, Alessandra Merenda, Sajith Perera, Tiago Grego, Jihoon Kim, Katie Andrews, Juergen Fink, William C. Skarnes, Bon-Kyoung Koo
bioRxiv 056549; doi: https://doi.org/10.1101/056549

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