Fig. 2: Development of GNGT2 MiniPromoters for cone cells.

A Bioinformatic design of Ple347. ENCODE segments that potentially regulate the expression of the GNGT2 gene are highlighted horizontally. Vertically highlighted genomic regions correspond to their color-matched segments included in the MiniPromoter design and are numbered as promoter(s) (P) and regulatory region(s) (RR). B Postnatal day 0 intravenous injection of Ple347-EmGFP, harvested 4 weeks later, led to robust expression in the outer nuclear layer (ONL). Scale bar, 100 µm. (For ubiquitous smCBA promoter see Fig. S1A). C Adult intravitreal injection of Ple347-EmGP and Ple349 (PDE6H)-EmGFP, harvested 4 weeks later, was quantified for epifluorescence intensity in opsin-positive cells, and showed that Ple347 (GNGT2) expression was significantly increased (~3×) compared to smCBA (p < 0.001). In addition, Ple349 (PDE6H) expression was significantly increased compared to both smCBA (~4×) (p < 0.001) and Ple347 (~1.5×) (p < 0.05). (For smCBA see Fig. S1D.) D Adult intravitreal injection of Ple347-EmGFP, harvested 4 weeks later, led to robust expression in cone cells, as indicated by co-staining with the cone cell marker opsin where EmGFP inner segments align to anti-opsin outer segments. Scale bar, 20 µm. (For smCBA see Fig. S1D.) EmGFP emerald green fluorescent protein, GCL ganglion cell layer, INL inner nuclear layer, IPL inner plexiform layer, N number of cells counted, OPL outer plexiform layer, TF transcription factor, TSS transcription start site. Green, anti-GFP; blue, Hoechst; red, anti-opsin.