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. 2019 Jan 15;14(1):e0208237. doi: 10.1371/journal.pone.0208237

Fig 4. Genotype-phenotype relationships in clonal BFU-E colonies derived from edited hematopoietic stem/progenitor cells.

Fig 4

A) Workflow depicting erythroid colony (BFU-E) formation genotype/phenotype assay using edited HSPC. HSPC were edited using RNPs targeting the desired region, plated in methocult, and colonies were grown for 14 days. Erythroid colonies were picked and genotyped by next-generation sequencing and phenotyped for their globin gene expression profile using RNA-seq. B) ß-globin (HBB) expression in erythroid colonies after gene editing, determined by RNA-seq. There was no statistically-significant difference in expression between each genotype of colony. C) 𝛾-globin (HBG1+HBG2) expression in erythroid colonies after gene editing, determined by RNA-seq. Because of the high variation between colonies of the same genotype, there was no statistically-significant difference in expression between each genotype of colony. To control for effects of RNP electroporation, the unedited cells were electroporated with a non-targeting RNP against BFP.