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- Title
High-resolution characterization of gene function using single-cell CRISPR tiling screen.
- Authors
Yang, Lu; Chan, Anthony K. N.; Miyashita, Kazuya; Delaney, Christopher D.; Wang, Xi; Li, Hongzhi; Pokharel, Sheela Pangeni; Li, Sandra; Li, Mingli; Xu, Xiaobao; Lu, Wei; Liu, Qiao; Mattson, Nicole; Chen, Kevin Yining; Wang, Jinhui; Yuan, Yate-Ching; Horne, David; Rosen, Steven T.; Soto-Feliciano, Yadira; Feng, Zhaohui
- Abstract
Identification of novel functional domains and characterization of detailed regulatory mechanisms in cancer-driving genes is critical for advanced cancer therapy. To date, CRISPR gene editing has primarily been applied to defining the role of individual genes. Recently, high-density mutagenesis via CRISPR tiling of gene-coding exons has been demonstrated to identify functional regions in genes. Furthermore, breakthroughs in combining CRISPR library screens with single-cell droplet RNA sequencing (sc-RNAseq) platforms have revealed the capacity to monitor gene expression changes upon genetic perturbations at single-cell resolution. Here, we present "sc-Tiling," which integrates a CRISPR gene-tiling screen with single-cell transcriptomic and protein structural analyses. Distinct from other reported single-cell CRISPR screens focused on observing gene function and gene-to-gene/enhancer-to-gene regulation, sc-Tiling enables the capacity to identify regulatory mechanisms within a gene-coding region that dictate gene activity and therapeutic response. Identifying functional domains and genetic regulatory mechanisms is essential for developing new therapies. Here the authors present sc-Tiling, single-cell high-density CRISPR tiling screening for functional domain characterization.
- Subjects
CRISPRS; RNA sequencing; GENOME editing; GENE expression; GENES; PROTEIN analysis
- Publication
Nature Communications, 2021, Vol 12, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-021-24324-0