B6 cg gt rosa 26sortm6 cag zsgreen1 hze j

Animal experiments were conducted in accordance with the Canadian Council for Animal Care guidelines and following approval from the University of Calgary Animal Care Committee or in accordance with National Institutes of Health and Mayo Foundation institutional guidelines. Six-to-8-week-old C57Bl/6, B6.Cg-Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}/J, and B6.Cg-Gt(ROSA)26Sor^{tm9(CAG-tdTomato)Hze}/J mice were purchased from Jackson Laboratories (Bar Harbor, ME, US). EoCRE mice were previously described^{27 (link)} and generously provided by Dr. J.J. Lee at the Mayo Clinic, Scottsdale. A key finding was that heterozygote mice expressed sufficient amounts of CRE recombinase to enable excision of targets expressed at the flox-stop-flox locus in ROSA26 while maintaining sufficient amounts of EPX^{27 (link)}. EoCRE^+/−/eGFP^+/− and eoCRE^+/−/tdTomato^+/− mice colonies were bred, housed, and maintained at the University of Calgary Animal Resource Centre. All imaging experiments were carried out at the Live Cell Imaging Laboratory at the University of Calgary.

Mouse genomic DNA samples obtained from tail snips were verified by PCR using primers Dhdds-FWD: 5′-GTGTCATCCCCTGCTGCAGAT-3′ and Dhdds-REV: 5′-TGGGTGTAGTG-GCTCAGGTC-3′ for genotype identification of floxed Dhdds alleles designed in a region which was conserved in both wild type (WT) and floxed alleles and also in the region flanking the loxP sites. The expected PCR product sizes for the WT and floxed alleles are 393 and 517 bp, respectively, thus differentiating WT, heterozygous floxed, and homozygous floxed alleles. PCR verification of Cre transgene modification was carried out using the following forward and reverse primer sets for Cre RPE 5′-AGGTGTAGAGAAGGCACTTAGC-3′ and 5′-CTAATCGCCATCT-TCCAGCAGG-3′, respectively, yielding a 411 bp product. RPE specific expression and activity of Cre-recombinase in VMD2-RPE Cre was verified by breeding these mouse lines against an ZsGreen reporter mouse strain (B6.Cg-Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}/J, Stock# 007906; The Jackson Laboratory, Bar Harbor, ME, USA) and monitoring ZsGreen expression in the retina.

HRGP^cre mice, which express Cre recombinase in cone photoreceptors, were described previously.¹⁴ (link) Ai6 (RCL-ZsGreen) mice [B6.Cg-Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)/Hze}/J], a Cre reporter line,¹⁵ (link) were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). Gjd2^fl/fl were generated as described in Yao et al.¹⁶ (link) and generously provided by David Paul, Harvard University. Mice were genotyped by PCR analysis of genomic DNA. Mice of both genders were used at 3 to 6 months of age. They were housed under a 12-hour light – 12 hours of dark cycle of illumination (luminance of 50-100 lux at cage level) with lights on from zeitgeber time (ZT) 0 to 12, and were given food and water ad libitum. All experimental procedures in “dark” conditions were performed under dim red light. Mice were euthanized by CO₂ asphyxiation followed by cervical dislocation. All procedures were approved by Emory University's Institutional Animal Care and Use Committee and conformed to the guidelines of the National Institutes of Health Guide for the Care and Use of Laboratory Animals and the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.