Intriguingly, the distribution patterns of the sgRNAs for SEMA6B, MT1E, and S100A4 - three of the four validated tumor suppressors-align well with the tumor pseudotime trajectory, suggesting the formation of diverse tumor genotypes in a single pooled screen that can recapitulate the whole epectrum of tumor evolution, from regions with fewer sgRNAs to colonies with substantial sgRNA enrichment (Figures 4E, 4F). Here, S100A4 is linked to neoplasm.