In vivo crispr/cas9 screening to simultaneously evaluate gene function in mouse skin and oral cavity

Sampath Kumar Loganathan, Ahmad Malik, Ellen Langille, Chen Luxenburg, Daniel Schramek

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article numbere61693
Pages (from-to)1-19
Number of pages19
JournalJournal of Visualized Experiments
Volume2020
Issue number165
DOIs
StatePublished - Nov 2020

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