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A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
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Chapitres
- 00:05Titre
- 00:50Cloning of gRNAs into the CRISPER-concatemer Vector
- 03:40Assessing Success of Cloning by Restriction Digest
- 05:28Preparing Intestinal Organoids for Electroporation
- 08:08Electroporation
- 10:36Results: Successful Generation of CRISPER-concatemers and Electroporation of Intestinal Organoids
- 11:24Conlcusion
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.
