Constructions of Minimal CRISPRa and CRISPRi Transgenes into pAAV Vector Backbone
To achieve efficient packaging of dSaCas9 fusion proteins and its sgRNA in a single AAV vector, a truncated version of the mouse Mecp2 brain-specific promoter (235 bp, pMecp2); a minimal polyadenylation signal (48 bp, spA); and the smallest version of the transcriptional regulator domain, such as VP64 activator (156 bp), VP160 activator (390 bp), KRAB repressor (227 bp), and SID4X repressor (432 bp), were used.