Talk Title:
In Situ Mapping of Cancer Mechanisms and Microenvironments via Pooled Perturbations

Abstract:

We developed CRISPRmap, a novel in situ barcode readout approach with concurrent multiplexed immunofluorescence and in situ RNA detection. CRISPRmap reduces both dependency on third party proprietary sequencing reagents and assay cost. We conducted a multi-omic base-editing screen in a breast cancer context on core DNA damage repair genes involved in the homologous recombination and Fanconi anemia pathways. Our approach enabled us to pinpoint likely-pathogenic patient-derived mutations that were previously classified as variants of unknown clinical significance. Furthermore, CRISPRmap effectively distinguished barcodes of a pooled library in tumor tissue, and we coupled it with cell-type and molecular phenotyping by cyclic immunofluorescence. Multi-omic spatial analysis of how CRISPR-perturbed cells respond to various environmental cues in the tissue context offers the potential to significantly expand our understanding of tissue biology in both health and disease.