The Application and Prospect of Single-cell CRISPR Screening Technology in Tumor Research

Abstract

Traditional population sequencing methods struggle to directly correlate gene perturbations with transcriptome changes at the single-cell level, due to limitations such as long cycle times and significant batch effects. Single-cell sequencing (SCS) can reveal the diversity and dynamics of cell populations, while CRISPR gene editing and screening technologies enable large-scale functional gene discovery. The combination of these two approaches has led to the development of ScCRISPR-seq. This article reviews the principles of CRISPR screening and SCS technologies, introduces the development of ScCRISPR-seq, and, drawing on research on lung cancer and intrahepatic cholangiocarcinoma (ICC), summarizes its key applications in cancer research, including deciphering tumor heterogeneity, elucidating mechanisms of drug resistance, and modulating the immune microenvironment. This article further explores challenges with this technology, including experimental workflow, data sparsity, and technical noise. Solutions are proposed, including data completion using AI algorithms, eliminating amplification bias using UMI tags, and optimizing gene-phenotype associations through network modeling. However, clinical translation is limited by unclear mechanisms of action and high barriers to entry in drug development. Future research may consider combining ScCRISPR-seq with network pharmacology and molecular docking techniques to improve the efficiency of new cancer drug development and advance precision immunotherapy.