Which cells should I use for genome-wide sgRNA screen?
Which cells should I use for genome-wide sgRNA screen?
The cells need to be a good surrogate for your experimental system but easy to grow and transduce. Primary cells might be the goal but can be difficult to scale up because they often do not proliferate or require elaborate culture conditions. We suggest substituting with a related transformed line for the primary screen, followed by use of more relevant primary cells for follow-up confirmation tests or more targeted screens.
Additionally, because we use lentivirus for delivery of the CRISPR components (Cas9 and sgRNA), it is wise to initially avoid cells that are hard to infect, i.e., those that require high MOI of VSV-g pseudotyped virus for efficient transduction.