CRISPR-Cas9-mediated insertion of a short artificial intron for the generation of conditional alleles in mice

Annelise Cassidy; Stephane Pelletier

doi:10.1016/j.xpro.2023.102116

CRISPR-Cas9-mediated insertion of a short artificial intron for the generation of conditional alleles in mice

STAR Protoc. 2023 Mar 17;4(1):102116. doi: 10.1016/j.xpro.2023.102116. Epub 2023 Feb 9.

Authors

Annelise Cassidy¹, Stephane Pelletier²

Affiliations

¹ Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN 46202, USA. Electronic address: amcassid@iu.edu.
² Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN 46202, USA. Electronic address: spellet@iu.edu.

Abstract

In this protocol, we describe the generation of conditional alleles in mice using the DECAI (DEgradation based on Cre-regulated Artificial Intron) approach. We detail steps for the CRISPR-mediated insertion of the short DECAI cassette within exon 3 of Scyl1 and the functional validation of alleles at genomic, transcriptomic, and protein levels. This strategy simplifies the process of generating mice with conditional alleles. For complete details on the use and execution of this protocol, please refer to Cassidy et al. (2022).¹.

Keywords: CRISPR; Developmental biology; Genetics; Model Organisms; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alleles
Animals
CRISPR-Cas Systems* / genetics
Exons / genetics
Gene Editing* / methods
Introns / genetics
Mice