Figure 5
From: Small molecules enhance CRISPR/Cas9-mediated homology-directed genome editing in primary cells
Effects of small molecules on the screening of knock-in porcine fetal fibroblasts cell lines. (A) Schematic diagram of the ROSA26 HDR template vector and strategy for insertion of the neomycin resistance gene into the pig ROSA26 intron 1 region. Red arrows are the binding sites of the primer sets to detect the insertion of the neomycin resistance gene to the targeted site. (B) Detection of knock-in cell lines using the primer sets shown in A. (C) Improvement of the percentage of knock-in cell lines in the presence of the three molecules in porcine fetal fibroblasts. Total represents the number of picked-up cell lines in each group, and Knock-in represents the number of knock-in cell lines identified by PCR and sequencing. (D) Representative sequencing results of junction regions of the inserted fragment at the targeted site in knock-in cell lines. Sequencing confirms precise insertion of neomycin resistance gene to targeted site of ROSA26 intron 1 region.
