CRISPR screening by AAV episome-sequencing (CrAAVe-seq) is a highly scalable cell type-specific <i>in vivo</i> screening platform.

ABSTRACT: There is a significant need for scalable CRISPR-based genetic screening methods that can be applied directly in mammalian tissues in vivo while enabling cell type-specific analysis. To address this, we developed an adeno-associated virus (AAV)-based CRISPR screening platform, CrAAVe-seq, that incorporates a Cre-sensitive sgRNA construct for pooled screening within targeted cell populations in mouse tissues. We demonstrate the utility of this approach by screening two distinct large sgRNA libraries, together targeting over 5,000 genes, in mouse brains to create a robust profile of neuron-essential genes. We validate two genes as strongly neuron-essential in both primary mouse neurons and in vivo, confirming the predictive power of our platform. By comparing results from indivi

SUBMITTER: Ramani B 

PROVIDER: S-EPMC10312723 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature