Supplementary Figure 1: Characterization of dCpf1 for gene activation.
From: Inducible and multiplex gene regulation using CRISPR–Cpf1-based transcription factors
(a) Schematic of doxycycline-inducible GFP reporter stably integrated into HEK293T cells used to assess CRISPRa activity. Promoter shown contains seven repeated activator target sites.
(b) Maps for expression constructs encoding dCas9, dLbCpf1, and dAsCpf1 fusions to the VP64-p65-Rta (VPR) activator.
(c) Quantification of GFP activation measured 3 days post-transfection with vectors expressing the protein fusions and guide RNAs. The rTTA+Dox serves as a positive control for GFP activation. Fluorescence values were normalized to HEK293-GFP reporter cells without plasmids transfected (column 1). Each bar is the mean of two replicates.
(d) Western blot analysis of HA-tagged dCas9, dLbCpf1, and dAsCpf1 fusions to VPR. The top band is the P2A-BFP attached to the fusion (P2A uncleaved) and the bottom band is the mature fusion without the P2A-BFP attached (P2A cleaved). The anti-GAPDH panel serves as a loading control.
