Supplementary Figure 2 : Analysis of various crRNAs targeted to the human CD5 and CD22 promoters.

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

Supplementary Figure 2

32 crRNAs (16 for each gene) were designed to target sites located within promoter sequences 1 kb upstream or 500 bps downstream of the TSS of each gene. Repetitive sequences were not targeted. After 72 hours post-transfection, cells were stained with fluorescently labeled antibody for (a) CD5 or (b) CD22 protein and fluorescent-positive cells were quantified by flow cytometry. Error bars represent s.e.m. for three biological independent replicates. Red circles indicate samples that are significantly different (Student t-test, two-tailed test assuming equal variance, p <0.05) compared to controls in which no crRNA was expressed. (c) and (d) are examples of experimental flow cytometry plots stained for CD5 and CD22 activation, respectively. (e) and (f) are examples of flow cytometry plots for negative controls stained for CD5 and CD22, respectively.