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A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
JoVE Journal
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A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer

A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer

17,592 Views

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11:53 min

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July 12, 2017

DOI:

10.3791/55916-v

DOI:
10.3791/55916-v

•

11:53 min
July 12, 2017

•

34 Views
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1Wellcome Trust – Medical Research Council Stem Cell Institute,University of Cambridge, 2Genetik Bölümü,University of Cambridge, 3Patoloji Bölümü,Yonsei University College of Medicine

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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.

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Merenda, A., Andersson-Rolf, A., Mustata, R. C., Li, T., Kim, H., Koo, B. A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer. J. Vis. Exp. (125), e55916, doi:10.3791/55916 (2017).

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