Products/Services Used | Details | Operation |
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Plasmid DNA Preparation> | … 2 Materials. 2.1 Cloning and PCR. 1. Synthesized oligo pool (eg, Genscript or Twist Bioscience). 2. lentiGuide-Puro sgRNA expression vector (Addgene, plasmid ID 52963). 3 … | Get A Quote |
Pooled CRISPR screens are emerging as a powerful tool to dissect regulatory networks, by assessing how a protein responds to genetic perturbations in a highly multiplexed manner. A large number of genes are perturbed in a cell population through genomic integration of one single-guide RNA (sgRNA) per cell. A subset of cells with the phenotype of interest can then be enriched through fluorescence-activated cell sorting (FACS). SgRNAs with altered abundance after phenotypic enrichment allow identification of genes that either promote or attenuate the investigated phenotype. Here we provide detailed guidelines on how to design and execute a pooled CRISPR screen to investigate molecular phenotypes. We describe how ... More