Fig. 6: Design of CRISPRa gRNAs for human gene MMP7 for different CRISPR nucleases.

a Schematic showing the steps involved in designing CRISPRa gRNAs targeting the promoter region of MMP7. A gene model and a list of CAGE peaks are used to define the optimal window for gene activation. A GuideSet is created separately for each CRISPR nuclease. DNase I hypersensitive site (DHS) information is obtained from AnnotationHub and added to the gRNA annotation. b The top track shows the promoter region of human gene MMP7 on chromosome 11, including part of the 5\({}^{\prime}\) UTR of MMP7 (yellow). The DHS and CAGE peak gray boxes were obtained using AnnotationHub (see Methods). The light pink region corresponds to the optimal region of activation, corresponding to a region [75,150]bp upstream of the \({5}^{\prime}\) end of the CAGE peak. For each of the four selected nucleases, all canonical PAM sites located within the optimal region are shown. PAM sites are colored by their on-target score: DeepHF for SpCas9, DeepCpf1 for AsCas12a, and enPAM+GB for enAsCas12a. No on-target scoring algorithm was available at time of publication for SpGCas9. The last track corresponds to common SNPs obtained from dbSNP151.