Journal
/
生物工程
/
A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
JoVE 杂志
生物工程
This content is Free Access.
JoVE 杂志 生物工程
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

•

11:53 min

•

July 12, 2017

DOI:

10.3791/55916-v

DOI:
10.3791/55916-v

•

11:53 min
July 12, 2017

•

34 Views
, , , , ,
1Wellcome Trust – Medical Research Council Stem Cell Institute,University of Cambridge, 2Department of Genetics,University of Cambridge, 3Department of Pathology,Yonsei University College of Medicine

Summary

Automatically generated

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.

Explore More Videos

CRISPR-concatemerMultiple Gene KnockoutMouse Small Intestinal OrganoidsCRISPRGuide RNAsElectroporationSimultaneous KnockoutLoss Of Function StudiesParallel CompensationCloningBbs1 Shuffling ReactionThree GRNA ConcatemerFour GRNA Concatemer

Read Article

Cite this Article

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).

Download .ris file

Copy

Share Video

.

Copy