Supplementary Figure 6: Exon-targeting 20-nt guides do not induce levels of transcriptional activation comparable to those of 15-nt and 20-nt guides targeted to promoter regions.
From: Multiplexed orthogonal genome editing and transcriptional activation by Cas12a
(a) Schematic of the approach to investigate the impact of 20-nt exon targeting guides on transcriptional activation. A representative gene model is shown; not drawn to scale. 15-nt as well as 20-nt guides targeting the promoter region or 20-nt guides targeting different genomic loci within exons were used. Three different qPCR primer pairs were used located either before or after the respective genomic loci of exon-targeting guides in order to measure transcriptional activation. (b) Relative mRNA expression after co-transfection of HEK293T cells with AsCas12a-VPR and crRNAs. 15-nt as well as 20-nt guides targeting the promoter region or 20-nt guides targeting different genomic loci within exons of TTN, NPY1R or MYOD1 were used. N = 2 independent experiments.
