Figure 4.

Interference of Endogenous Gene Expression with DCUg

Cells were transfected by various vectors and detected at 24 h post transfection. (A) Flow cytometry analysis of cell apoptosis. (B) Expression level of Cas13a mRNA in HepG2 and HL7702 cells. (C) qPCR analysis of mRNA expression. (D) Western blot analysis of protein expression. The representative image and quantified optical density were shown. Lip, Lipofectamine; NT/TERT/EZH2/RelA/TER, pDCUg-NT/TERT/EZH2/RelA/TER. (E) CCK-8 assay of cell growth. Blank, cells without any treatment. Lipofectamine, cells treated with Lipofectamine only. pDCUg-NT/TERT/EZH2/RelA/TER, plasmid expressing DMP-controlled Cas13a and U6-controlled gRNA targeting no transcript (NT), TERT, EZH2, RelA, and TER transcripts. TER, TERT&EZH2&RelA. All values are mean ± SEM with n = 3, unless otherwise noted. ∗p < 0.05; ∗∗p < 0.01.