Creating pooled CRISPR-Cas9 knock-outs in NIH-3T3 cells
Output Details
Description
To validate a genome wide CRISPR screen, we select the top hits and create lentiviruses to validate the hits. Rather than screening each virus from single cell clones, we analyze the infected cells as pools according to the following protocol. This method involves three major steps: Create Cas9-expressing 3T3 cells (1-3 months) Make lentiviruses that carry sgRNA (2 weeks) Infect 3T3-Cas9 cells and assess gRNA efficiency (1 week)
Identifier (DOI)
10.17504/protocols.io.eq2ly7wpmlx9/v1
