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Pooled In Vitro and In Vivo CRISPR-Cas9 Screening Identifies Tumor Suppressors in Human Colon Organoids
Highlights • An optimized protocol for pooled CRISPR-Cas9 library screening in human colon organoids • Organoid xenografts enable unbiased identification of in vivo tumor suppressors • gRNA functionality in organoids is less predictable compared to 2D cancer cell lines • Clonal tracing with a UMI library allows adjustment for clonal drift during selection Summary Colorectal cancer (CRC) is characterized by prominent genetic and phenotypic heterogeneity between patients.
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