Fig. 2
From: High throughput single-cell detection of multiplex CRISPR-edited gene modifications
a Schema of generation of a multiplexed Ba/F3 cell line. b CRISPR-seq NGS validating the presence of gene edits for all 6 targeted loci. c Workflow for single-cell qPCR detection of sgRNAs by Fluidigm Biomark. Single cells from the Ba/F3 multiplexed line were sorted (based on viable, DAPI−, and transduced mCherry+) into one PCR plate, then analyzed using the Biomark assay. d Histogram reporting the number of on-target gene edits detected across 3429 single cells, from the multiplexed-edited Ba/F3 cell line. e Modification rates across single cells containing at least one gene edit (rows) for the six on-target loci (columns) using single-cell amplicon sequencing. Cells (n = 3200 pre-transplant, n = 2747 post-transplant) were assayed before and after in vivo passaging, obtained via intravenous injection into NSG mice. Splenocytes were sampled at euthanasia. A scale bar ranging from 0% (blue color) to 100% (red color) modifications is shown. f Histogram showing the number of cells containing mutations for the indicated gene combinations. Set size refers to the number of cells for each of the six targets. Intersection size refers to the number of cells for each gene-edit combinatorial assortment. g Combinatorial assortments of cells post-transplant