A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
Journal of Visualized Experiments
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Merenda, A., Andersson-Rolf, A., Mustata, R., Li, T., Kim, H., & Koo, B. (2017). A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer. Journal of Visualized Experiments, (125)https://doi.org/10.3791/55916
This protocol describes the steps for cloning multiple single guide RNAs into one guide RNA concatemer vector, which is of particular use in creating multi-gene knockouts using CRISPR/Cas9 technology. The generation of double knockouts in intestinal organoids is shown as a possible application of this method.
A.M. is supported by Wntsapp (Marie Curie ITN), A.A-R. is supported by the Medical Research Council (MRC), and B-K.K. and R.M. are supported by a Sir Henry Dale Fellowship from the Wellcome Trust and the Royal Society [101241/Z/13/Z] and receive support through a core grant from the Wellcome Trust and MRC to the Wellcome Trust - MRC Cambridge Stem Cell Institute.
Wellcome Trust (101241/Z/13/Z)
External DOI: https://doi.org/10.3791/55916
This record's URL: https://www.repository.cam.ac.uk/handle/1810/265699