In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity.
J Vis Exp
; (165)2020 11 02.
Article
en En
| MEDLINE
| ID: mdl-33191932
ABSTRACT
Genetically modified mouse models (GEMM) have been instrumental in assessing gene function, modeling human diseases, and serving as preclinical model to assess therapeutic avenues. However, their time-, labor- and cost-intensive nature limits their utility for systematic analysis of gene function. Recent advances in genome-editing technologies overcome those limitations and allow for the rapid generation of specific gene perturbations directly within specific mouse organs in a multiplexed and rapid manner. Here, we describe a CRISPR/Cas9-based method (Clustered Regularly Interspaced Short Palindromic Repeats) to generate thousands of gene knock-out clones within the epithelium of the skin and oral cavity of mice, and provide a protocol detailing the steps necessary to perform a direct in vivo CRISPR screen for tumor suppressor genes. This approach can be applied to other organs or other CRISPR/Cas9 technologies such as CRISPR-activation or CRISPR-inactivation to study the biological function of genes during tissue homeostasis or in various disease settings.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Piel
/
Pruebas Genéticas
/
Sistemas CRISPR-Cas
/
Proteína 9 Asociada a CRISPR
/
Boca
Tipo de estudio:
Diagnostic_studies
/
Guideline
/
Prognostic_studies
/
Screening_studies
Límite:
Animals
Idioma:
En
Revista:
J Vis Exp
Año:
2020
Tipo del documento:
Article