Unlike pooled and arrayed CRISPR libraries for gene knockout, each EdiGene’s Edi-G knockout (KO) cell line is individually cloned and validated by Sanger sequencing. The pathway or cancer-related Edi-G KO cell lines are provided with parental wild type cell line(s) for direct functional and phenotypic comparison. A direct relationship among signaling pathway or cancer-related genes could be established or confirmed if one gene’s loss-of-function results in over expression or loss-of-function of other gene(s). Because these cell line pairs differ only by the knockout of a specific gene but share the same cellular background, the biological impact and gene-gene relationship could be unambiguously established, without the typical confirmation steps needed for pooled or arrayed CRISPR knockout.