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. 2018 Jul 10;4:36. doi: 10.1038/s41421-018-0035-0

Fig. 2. catRNA enhances knock-out and knock-in in mammalian cells.

Fig. 2

a Schematic flowchart of the catRNA maturation process and subsequent gene disruption. b Targeted gene knock-out with crRNA, caRNA, and catRNA in 293T cells. Gene knock-out was performed by co-transfecting the LbCpf1 DNA plasmid and plasmids encoding different crRNA species with Lipofectamine 3000. Specific gene knock-outs and knock-ins were measured 3 days after transfection with TIDE online software. c Targeted knock-in with crRNA, caRNA, and catRNA in 293T cells. Knock-in was performed with LbCpf1 and different crRNA species together with ssDNA donor templates. An additive effect was always observed when the 5′ cap, 3′ tail, and truncated tRNA structures were used in combination in knock-outs and knock-ins with catRNA. Specific gene knock-outs and knock-ins were measured 3 days after transfection with TIDE online software. Bar, SE. n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 determined by the Mann–Whitney test