Skip to main content
. 2022 Sep 7;2(9):100290. doi: 10.1016/j.crmeth.2022.100290

Figure 1.

Figure 1

Cas9 modRNA and sgRNA efficiently knock out an integrated GFP in hPSCs

(A) Schematic diagram for knocking out GFP in H1 OCT4-GFP cells using Cas9 modRNA and in vitro-synthesized sgRNA.

(B) DNA templates used to synthesize modRNA for Cas9-2A-GFP, Cas9, Cas9-2A-Puro, p53DD, and ABE8e, including a summary of all the constructs used in this work.

(C) H1 OCT4-GFP cells were cultured on iMatrix-511 in mTeSR1 and transfected with different combinations of Cas9 modRNA and GFP sgRNA. On day 4, cells were collected, and GFP expression was analyzed via flow cytometry. The percentage of GFP− cells for each combination is shown in the form of a tiled heatmap. Experiments were repeated three times, and representative data are shown.

See also Figure S1.