Skip to main content
. Author manuscript; available in PMC: 2019 Jan 9.
Published in final edited form as: Cell Rep. 2018 Dec 18;25(12):3262–3272.e3. doi: 10.1016/j.celrep.2018.11.079

Figure 3. Inhibitory Effects of PS-Modified DNA Oligonucleotides with Random Sequences on Cpf1-Mediated Genome Editing in Human Cells.

Figure 3.

(A). Inhibitory effects of ps42DNA-AAVS1 and ps42DNA-FANCF on AsCpf1-mediated genome editing at the DNMT1 locus in 293T cells. ps42DNA-AAVS1 or ps42DNA-FANCF was added simultaneously with AsCpf1 mRNA and AsCpf1 crRNA targeting DNMT1 locus.

(B). Inhibitory effects of RAMpsDNA on AsCpf1-mediated genome editing at the DNMT1, AAVS1, and FANCF loci in 293T cells. RAMpsDNA was added simultaneously with CRISPR-Cpf1 mRNA and crRNA.

(C).Inhibitory activity of RAMpsDNA on AsCpf1-mediated genome editing at the DNMT1 locus in Hep3B cells. RAMpsDNA was added simultaneously with CRISPR-Cpf1 mRNA and crRNA targeting DNMT1 locus.

(D). Time-dependent inhibitory effects of RAMpsDNA on AsCpf1-mediated genome editing at the DNMT1 locus in 293T cells. ‘‘Time to add inhibitor’’: RAMpsDNA was added at various time points after treatment with AsCpf1 mRNA and crRNA targeting DNMT1 locus.

The control group (Ctrl) was treated only with AsCpf1 mRNA and crRNA. The concentration of inhibitors is 140 nM (2.5-fold molar excess relative to crRNA). Relative genome-editing efficiency in (A)–(D) was determined using the T7E1 cleavage assay from three biological replicates 48 hr post-treatment and normalized to that of the control group without adding inhibitors. ***p < 0.001 versus the control group, two-tailed t test. See Figure S3 for the corresponding gel images.