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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: Biotechnol Bioeng. 2019 Nov 12;117(2):593–598. doi: 10.1002/bit.27199

Figure 1. Components for CRISPRa.

Figure 1

A) gRNAs were designed using the CHO-K1 annotation from NCBI. Since experimentally reported TSSs have not been reported for CHO cells, the TSS was defined as the starting base pair of the annotated target genes. Three gRNAs (red rectangles) were designed between 25 bp and 550 bp upstream of the TSS. gRNA positions for each gene targeted in this study are given in Supplementary Table S2. B) A plasmid encoding VPR-dCas9 was transfected along with either one of the gRNA plasmids or a mix of all gRNA plasmids. The gRNAs guide the VPR-dCas9 to a region upstream of the TSS where VPR increases expression of the target gene.