Arrayed CRISPR/Cas9 Screening for the Functional Validation of Cancer Genetic Dependencies
L Proietti, G Manhart, E Heyes, S Troester, F Grebien - Bio-protocol, 2022 - bio-protocol.org
CRISPR/Cas9 screening has revolutionized functional genomics in biomedical research
and is a widely used approach for the identification of genetic dependencies in cancer cells.
Here, we present an efficient and versatile protocol for the cloning of guide RNAs (gRNA)
into lentiviral vectors, the production of lentiviral supernatants, and the transduction of target
cells in a 96-well format. To assess the effect of gene knockouts on cellular fitness, we
describe a competition-based cell proliferation assay using flow cytometry, enabling the …
and is a widely used approach for the identification of genetic dependencies in cancer cells.
Here, we present an efficient and versatile protocol for the cloning of guide RNAs (gRNA)
into lentiviral vectors, the production of lentiviral supernatants, and the transduction of target
cells in a 96-well format. To assess the effect of gene knockouts on cellular fitness, we
describe a competition-based cell proliferation assay using flow cytometry, enabling the …
