Figure 6.

Targeted Transgene Activation in the Brain Tissue by Systemic Delivery of AAV1-PHP.B Expressing CRISPRa

(A) pSyn1-iRFP720-GFP construct. The red arrows (34, 35, and 36) on the hSyn1 promoter indicate the locations of three different sgRNA target sites for dSaCas9-VP64. (B) Example of in vivo imaging of the mouse brains. AAV1-WT (left) or AAV1-PHP.B (right) expressing iRFP720 and GFP fluorescent proteins was intravenously injected into the mice. In vivo imaging was carried out 5 weeks after injection. iRFP720 signal was detected only at the head of the AAV1-PHP.B-injected mouse. (C) Ex vivo imaging of the dissected ventral brain tissues. AAV1-PHP.B expressing dSaCas9-VP64 was injected together with AAV1-PHP.B expressing iRFP720 and GFP fluorescent proteins. After seven weeks of AAVs injection, the mouse brains were harvested. Ex vivo imaging was then carried out to detect and quantify the GFP (left) and iRFP720 (right) fluorescent signals on the mouse ventral brains. The statistical significance levels from control (uninjected brain) are indicated as *p < 0.05, **p < 0.01, and ***p < 0.001. All data are presented as mean ± SD.