Direct measurement of engineered cancer mutations and their transcriptional phenotypes in single cells.

ABSTRACT: Genome sequencing studies have identified numerous cancer mutations across a wide spectrum of tumor types, but determining the phenotypic consequence of these mutations remains a challenge. Here, we developed a high-throughput, multiplexed single-cell technology called TISCC-seq to engineer predesignated mutations in cells using CRISPR base editors, directly delineate their genotype among individual cells and determine each mutation's transcriptional phenotype. Long-read sequencing of the target gene's transcript identifies the engineered mutations, and the transcriptome profile from the same set of cells is simultaneously analyzed by short-read sequencing. Through integration, we determine the mutations' genotype and expression phenotype at single-cell resolution. Using cell lines, we eng

SUBMITTER: Kim HS 

PROVIDER: S-EPMC11324510 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature