I intend to do CRISPR screening on a cell line made in HEK293T cells and I do have Whole Genome CRISPR library (Brunello) plasmid .; this plasmid has 17441 SgRNA targeting almost 900 non essential genes but it doesn't have cas9 . Can I make viral particles with these plasmid and use for CRIPSR screening without co infection of cas9 plasmids. Does HEK293T cells(Wild type) have Cas9 enough to run the CRISPR screening?
John Hardy Lockhart
Cas9 is a bacterial protein and is not expressed by eukaryotic cells. Since your library does not include the Cas9 protein on the same plasmid, you will need to perform an additional infection.
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