Figure S4. Exosome-mediated delivery of in vitro transcribed (IVT) synthetic guide RNA (sgRNA) induces gene editing in Cas9 overexpressing cells to disrupt oncogenic KrasG12D.
(A) Generation of IVT sgRNA. Starting from a plasmid sgRNA vector with T7 promoter, KrasG12D sgRNA1/2 was first PCR-amplified, run on agarose gel to confirm amplicon size, and DNA concentration was quantified with NanoDrop. The PCR products were purified and used as templates for the vitro synthesis of sgRNA. (B) HEK293T exosomes were loaded with IVT-KrasG12D sgRNA1 using Exo-Fect and used to treat Cas9-overexpressing KPC689 cells. Gene editing was tested with T7/Surveyor assay. (C) mRNA expression level of KrasG12D in Cas9-overexpressing cells was assessed by Quantitative PCR. Data are normalized to 18s and untreated control and presented as mean ± standard deviation. Unpaired t test was used to evaluate mean differences based on ΔCT values. ***P < 0.001.
Source data are available for this figure.
