High-resolution characterization of gene function using single-cell CRISPR tiling screen.

IF: 17.694

Cited by: 1

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.

Keywords

Seurat

Gene Expression

MeSH terms

CRISPR-Cas Systems

Clustered Regularly Interspaced Short Palindromic Repeats

Drug Screening Assays, Antitumor

Gene Editing

Gene Expression Regulation, Neoplastic

Genetic Testing

Genome, Human

Histone-Lysine N-Methyltransferase

Histones

Humans

Models, Molecular

Mutagenesis

Neoplasms

Phenotype

Transcriptome

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

Hoshii, Takayuki

Xiao, Gang

Müschen, Markus

Chen, Jianjun

Armstrong, Scott A

Chen, Chun-Wei

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