CRISPR Pooled Libraries

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Cartoon of four cells, with a different plasmid in each. A zoom panel shows the interior of the cells, where Cas9 uses a library sgRNA to cut a matching genomic target. Each plasmid has a different genomic target. An arrow labeled “screen for different outcomes” points to the four cells with the target genes knocked out, indicated with an X. Three cells are gray while one remains the same color, representing a different phenotype after knockout.
A cartoon of four cells, all carrying a different plasmid. Each cell glows a different color, matching the plasmid it carries.
Cartoon depicting factors to consider for a pooled library when creating an amplification protocol; details are in text of link.
Graphic showing flow chart of pooled library preparation.
Doench Libraries blog graphic
Summary of the advantages of the new approach to genome-wide CRISPR screen design: Safe-targeting guides better control for dsDNA break toxicity by targeting Cas9 to unannotated genomic regions. Off-target cutting doesn't confound screen results. Short gRNAs have lower off-target cuts than full-length gRNAs.
CRISPR Kinome Libraries
Strategy for assembling barcoded combinatorial gRNA libraries.

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