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Pooled Genome-Scale CRISPR Screens in Single Cells.
- Source :
-
Annual review of genetics [Annu Rev Genet] 2023 Nov 27; Vol. 57, pp. 223-244. Date of Electronic Publication: 2023 Aug 10. - Publication Year :
- 2023
-
Abstract
- Assigning functions to genes and learning how to control their expression are part of the foundation of cell biology and therapeutic development. An efficient and unbiased method to accomplish this is genetic screening, which historically required laborious clone generation and phenotyping and is still limited by scale today. The rapid technological progress on modulating gene function with CRISPR-Cas and measuring it in individual cells has now relaxed the major experimental constraints and enabled pooled screening with complex readouts from single cells. Here, we review the principles and practical considerations for pooled single-cell CRISPR screening. We discuss perturbation strategies, experimental model systems, matching the perturbation to the individual cells, reading out cell phenotypes, and data analysis. Our focus is on single-cell RNA sequencing and cell sorting-based readouts, including image-enabled cell sorting. We expect this transformative approach to fuel biomedical research for the next several decades.
- Subjects :
- Genetic Testing methods
Phenotype
CRISPR-Cas Systems genetics
Genome genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1545-2948
- Volume :
- 57
- Database :
- MEDLINE
- Journal :
- Annual review of genetics
- Publication Type :
- Academic Journal
- Accession number :
- 37562410
- Full Text :
- https://doi.org/10.1146/annurev-genet-072920-013842