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

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 a...

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Detalles Bibliográficos
Autores principales: Cassidy, Annelise, Pelletier, Stephane
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9943854/
https://www.ncbi.nlm.nih.gov/pubmed/36853660
http://dx.doi.org/10.1016/j.xpro.2023.102116
Descripción
Sumario: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)

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