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[Preprint]. 2023 Jan 20:2023.01.20.23284646. [Version 1] doi: 10.1101/2023.01.20.23284646

Figure 5. WT and mutant PAM promoter activity.

Figure 5.

A 5 kb PAM promoter-5’UTR WT sequence was cloned upstream of a luciferase reporter. Six mutant constructs were created by site-directed mutagenesis. The constructs were transiently transfected into HEK-293 AD cells together with a Renilla luciferase reporter for normalization. Firefly and Renilla luciferase activities were measured 24 h post-transfection. Two SNVs, c.−361G>A and c.−133T>C, have significantly lower transcriptional activity than the WT hybrid transcript. Mock, empty pRMT-Luc vector. Differences between experimental groups were analyzed by 1-way ANOVA with Dunnett’s post hoc test, using WT as the control group. RLU, relative luciferase activity. *, p = 0.0443; **, p = 0.0019; ***, p = 0.0005.