Spatially Resolved <i>in vivo</i> CRISPR Screen Sequencing via Perturb-DBiT.

ABSTRACT: Perturb-seq enabled the profiling of transcriptional effects of genetic perturbations in single cells but lacks the ability to examine the impact on tissue environments. We present Perturb-DBiT for simultaneous co-sequencing of spatial transcriptome and guide RNAs (gRNAs) on the same tissue section for in vivo CRISPR screen with genome-scale gRNA libraries, offering a comprehensive understanding of how genetic modifications affect cellular behavior and tissue architecture. This platform supports a variety of delivery vectors, gRNA library sizes, and tissue preparations, along with two distinct gRNA capture methods, making it adaptable to a wide range of experimental setups. In applying Perturb-DBiT, we conducted un-biased knockouts of tens of genes or at genome-wide scale across three canc

SUBMITTER: Baysoy A 

PROVIDER: S-EPMC11601513 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature