Figure 4.

Biological applications of scCRISPR. A) Dissecting molecular regulatory mechanisms and interactions. scCRISPR is performed to uncover interactions between regulator genes and those genes involved in unfolded protein response (UPR). CRISPRi of IRF8 blocked TFs interaction, preventing AP‐1 factor activity from being coordinated with that of IKZF1. TAP‐seq was developed to map 1778 enhancers to corresponding genes. In CRISPR‐sciATAC, the effect of distinct perturbations of chromatin modifiers and remodelers on chromatin accessibility was demonstrated. B) scCRISPR offers a profound understanding of tumor biology. scCRISPR technologies are promising and potent tools for investigating tumor drug resistance mechanisms via gene perturbation at scale. Through genetic screening, the tumor microenvironment and markers can be characterized, and the metastasis dynamics of cancer cells can be monitored using Perturb‐map. C) Spatially resolved genotype‐to‐phenotype mapping. Imaging‐based scCRISPR connects perturbed genes to optical cellular phenotypes, including protein translocation, stress granule, the functional structure of a protein, and organic shapes in zebrafish. D) Investigating the pathogenesis of the development of the neural system. In vivo Perturb‐seq redefines gene functions and identifies the genetic mechanism of ASD/ND in the murine‐developing brain.