Figure 2.

SynNotch-Cas9:p300 systems edit and activate immune genes with 20 bp- and 14–15 bp-length sgRNAs, respectively. (A) Diagram of the synNotch-Cas9:p300 system. (B and C) Editing of PD-1 and CTLA4 (B) and activation of CCL19 and IL2 (C) with the synNotch-Cas9:p300 system (H1) was detected through T7E1 and q-PCR assays, respectively. The numbered blue boxes indicate the binding regions of the 14–15 bp sgRNAs. (D and E) Editing of PD-1 and CTLA4 (D) and activation of CCL19 and IL2 (E) with the synNotch-Tet-Cas9:p300 system (eZ3) was examined. For the q-PCR: the results of one representative experiment out of three repeats are shown. Student’s t test, **P < 0.01, error bars, SEM. N.D., not detected. P.C., Jurkat cells transfected with plasmids encoding a constitutively expressed Cas9:p300 and the corresponding gRNAs. Blank, Jurkat cells