Supplementary Figure 10: Targeted LmoCascade editing can be achieved by tethering I-TevI monomeric endonuclease domains to Cas6. | Nature Biotechnology

Supplementary Figure 10: Targeted LmoCascade editing can be achieved by tethering I-TevI monomeric endonuclease domains to Cas6.

From: Targeted transcriptional modulation with type I CRISPR–Cas systems in human cells

Supplementary Figure 10

(a) Schematic of the IL1RN locus along with IL1RN crRNA target sites. (b) I-TevI-Cas6 effectors of different lengths driven by human cytomegalovirus (CMV) promoter. (c) Quantification of percent genomic DNA editing modifications generated following co-transfection of individual crRNAs and LmoCascade with I-TevI(184)-Cas6 or I-TevI(206)-Cas6 fusions in HEK293T cells. (d) Schematic of genomic DNA modifications at IL1RN cr14 target site and representative sequences of mutated alleles identified from deep sequencing analysis of HEK293T bulk cell population co-transfected with LmoCascade and I-TevI(184)-Cas6. I-TevI 5’-CNNNG-3’ nuclease motifs highlighted in orange. All samples processed at 4 days post transfection. (n=1 biological independent sample). TSS, transcription start site; NT, No treatment.

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