One-step generation of conditional and reversible gene knockouts
Andersson Rolf, Amanda
Nature Publishing Group
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Andersson Rolf, A., Mustata, R., Merenda, A., Kim, J., Perera, S., Grego, T., Andrews, K., et al. (2017). One-step generation of conditional and reversible gene knockouts. Nature Methods https://doi.org/10.1038/nmeth.4156
Loss-of-function studies are key for investigating 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 biallelic conditional gene knockouts in diploid or aneuploid cells, such as pluripotent stem cells, 3D organoids and cell lines, by co-delivery of CRISPR-Cas9 and a universal conditional intronic cassette.
A.A.-R. and K.T. are supported by the Medical Research Council, A.M. is supported by Wntsapp, Marie Curie ITN. J.F. and J.C.R.S. are supported by the Wellcome Trust. W.C.S. received core grant support from the Wellcome Trust to the Wellcome Trust Sanger Institute. B.-K.K. and R.C.M. are supported by a Sir Henry Dale Fellowship from the Wellcome Trust and the Royal Society (101241/Z/13/Z) and receive a core support grant from the Wellcome Trust and MRC to the WT–MRC Cambridge Stem Cell Institute.
Wellcome Trust (101861/Z/13/Z)
Wellcome Trust (097922/B/11/Z)
Wellcome Trust (101241/Z/13/Z)
External DOI: https://doi.org/10.1038/nmeth.4156
This record's URL: https://www.repository.cam.ac.uk/handle/1810/263063