Fig. 4 | Nature Communications

Fig. 4

From: A thermostable Cas9 with increased lifetime in human plasma

Fig. 4

Genome-editing activity of GeoCas9 in mammalian cells. a EGFP disruption in HEK293T cells by GeoCas9. HEK293FT cells expressing a destabilized GFP were transfected with GeoCas9 RNP preassembled with a targeting or non-targeting guide RNA. Cells were analyzed by flow cytometry and targets adjacent to the CRAA PAM resulted in efficient GFP disruption. b T7E1 analysis of indels produced at the AAVS1 locus when the guide length was varied from 21 to 22 nt. The Cas9 used is indicated above each lane and the length of the spacer is shown below. c T7E1 analysis of indels produced using a titration of GeoCas9 and SpyCas9 RNP targeting the DNMT1 locus in HEK293T cells. The Cas9 used is indicated above each lane and the amount of RNP delivered to each well of a 96-well plate is indicated below. NT non-targeting

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