Issue link: http://life-technologies.uberflip.com/i/438554
52 Life Technologies ™ | Genome editing GeneArt ® Genomic Cleavage Detection Kit 640 500 400 300 200 TALENs CRISPR TALENs CRISPR Lipofectamine ® 2000 Lipofectamine ® 3000 5 10 % Gene modification efficiency 0 B 5 10 15 20 % Gene modification efficiency 0 TALENs CRISPR TALENs CRISPR Lipofectamine ® 2000 Lipofectamine ® 3000 bp bp 500 400 300 200 100 A pcDNA™ 3.3 control pcDNA™ 3.3 control Figure 14. Cleavage efficiency of GeneArt ® TALs and CRISPRs. The TALs and CRISPRs targeted the AAVS1 locus in (A) U2OS cells, derived from human bone osteosarcoma, and (B) HepG2 cells, derived from a human hepatocellular carcinoma, after transfection with either Lipofectamine ® 2000 or 3000 reagent. Cleavage was assayed using the GeneArt ® Genomic Cleavage Detection Kit. Both cell lines showed improved transfection efficiency and protein expression compared to transfection using Lipofectamine ® 2000 reagent. Transfection efficiency and protein expression were assessed using a CRISPR construct that contains the orange fluorescent protein (OFP) reporter gene. We observed increased TAL- and CRISPR-mediated cleavage for the AAVS1 target locus in both cell lines transfected with Lipofectamine ® 3000 reagent, demonstrating that increasing the transfection efficiency and, by implication, protein expression, will increase the cleavage rate of TALs and CRISPRs. U2OS cells transfected with Lipofectamine ® 3000 reagent showed 1.5-fold improved TAL cleavage efficiency and slightly improved CRISPR cleavage (A). HepG2 cells had 3-fold higher cleavage efficiency for TAL effectors and 8-fold higher for CRISPRs (B).