Supplementary Figure 6 : Inducibility and reversibility of A/C heterodimerizer drug-regulated dLbCpf1-based activators.

From: Inducible and multiplex gene regulation using CRISPR–Cpf1-based transcription factors

Supplementary Figure 6

(a) To measure the kinetics of activator induction, HEK293 cells were transfected with plasmids expressing dLbCpf1-DmrA(x4), DmrC-p65, and MST crRNAs targeting the human HBB or AR promoters (same crRNAs used in Fig. 2b). 34 hours after transfection, these cells were split into two cultures: one with media containing A/C heterodimerizer (500uM) (top, blue) and one with media lacking the A/C heterodimerizer (bottom, purple). Cells were collected at various time points and relative mRNA expression levels were measured by RT-qPCR compared to a negative control. (b) To measure the kinetics of reversibility, HEK293 cells were transfected as in (a). 24 hours after transfection, A/C heterodimerzer (500uM) was added to the medium. 10 hours later, these cells were split into two cultures: one with media containing A/C heterodimerizer (500uM) (top, blue) and one with media lacking the A/C heterodimerizer (bottom, purple). Cells were collected at various time points and relative mRNA expression levels were measured by RT-qPCR compared to a negative control. Error bars represent s.e.m. of three biological independent replicates.