Rhodopsin mRNA expression levels.
a. Contralateral eyes of adult P347S mice (n=6) expressing a mutant human RHO transgene were subretinally injected with 6.0×108 vp AAV-S or AAV-R. Two weeks post-injection retinas were harvested, transduced (green) cells collected by FACS and RNA extracted. The level of human RHO suppression by AAV-S, determined by qPCR using human-specific RHO primers, was 68±2.4%. b. Adult wild type mice (n=7) were subretinally injected with a mixture of either 6.0×108 vp AAV-S and 1.8×1010 vp AAV-R or 6.0×108 vp AAV-C and 1.8×1010 vp AAV-R. Two weeks post-injection total RNA was extracted and levels of replacement RHO RNA determined by qPCR. No significant difference in replacement transcript levels was observed in eyes that received AAV-R and AAV-S versus AAV-R and AAV-C, indicating that no significant suppression of the replacement transcript had occurred (p=0.814). c. Adult wild type mice (n=8) were subretinally injected with AAV-R. Total RNA was extracted two weeks post-injection and levels of RHO replacement expression from AAV-R compared to levels of RHO in NHR mouse retinas. Levels of RHO mRNA expression from the AAV-R was 31±5% of levels in NHR mouse retinas.