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Gene knockout screens using pooled sgRNA libraries. A, schematic of forward genetic screens in human cells using pooled sgRNA libraries. B, crystal violet staining of 6-TG-resistant cells transfected previously with the sgRNA library. 6-TG-resistant cells appeared when sgRNAs targeting HPRT1 were diluted in the library containing 30,840 sgRNAs. The mutated sequences from surviving clones are shown below. Red letters indicate the PAM, and the blue letter indicates an insertion.
