Subcellular protein localizations and GFP integration sites in GFP-tagged clones isolated from the pool of tagged cells. (A) Representative images of individual clones isolated by single-cell dilution and identified by massively parallel sgRNA sequencing. Scale bars, 25 µm. (B) Comparison of localizations of 335 individually isolated clones to localization annotations in The Human Protein Atlas. (C) Outline of integration site analysis in a subpool of approximately 50 different GFP-tagged cells. (D) The 50-bp region upstream of the integration site aligns either to an on- or off-target genomic region or to the plasmid backbone in case the donor was only cut once, leading to integration of the whole donor plasmid, or to the 3′ end of an additional GFP fragment in case of a double integration event. (E) Integration sites and gene names of sgRNAs identified in the subpool by massively parallel sequencing (ranked by abundance). (F) Alignment of the identified integration sites in the subpool.
