Back to Search
Start Over
High-resolution characterization of gene function using single-cell CRISPR tiling screen.
- Source :
-
Nature communications [Nat Commun] 2021 Jul 01; Vol. 12 (1), pp. 4063. Date of Electronic Publication: 2021 Jul 01. - Publication Year :
- 2021
-
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.
- Subjects :
- Drug Screening Assays, Antitumor
Gene Editing
Gene Expression Regulation, Neoplastic
Genetic Testing
Genome, Human
Histone-Lysine N-Methyltransferase chemistry
Histone-Lysine N-Methyltransferase genetics
Histones
Humans
Models, Molecular
Mutagenesis
Transcriptome
CRISPR-Cas Systems
Clustered Regularly Interspaced Short Palindromic Repeats genetics
Neoplasms genetics
Phenotype
Subjects
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 12
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 34210975
- Full Text :
- https://doi.org/10.1038/s41467-021-24324-0