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| Content Provider | Springer Nature Link |
|---|---|
| Author | Casco Robles, Martin Miguel Miura, Tomoya Chiba, Chikafumi |
| Copyright Year | 2014 |
| Abstract | The adult newt has the ability to regenerate the neural retina following injury, a process achieved primarily by the retinal pigment epithelium (RPE). To deliver exogenous genes to the RPE for genetic manipulation of regenerative events, we isolated the newt RPE65 promoter region by genome walking. First, we cloned the 2.8 kb RPE65 promoter from the newt, Cynops pyrrhogaster. Sequence analysis revealed several conserved regulatory elements described previously in mouse and human RPE65 promoters. Second, having previously established an I-SceI-mediated transgenic protocol for the newt, we used it here to examine the −657 bp proximal promoter of RPE65. The promoter assay used with F0 transgenic newts confirmed transgene expression of mCherry fluorescent protein in the RPE. Using bioinformatic tools and the TRANSFAC database, we identified a 340 bp CpG island located between −635 and −296 bp in the promoter; this region contains response elements for the microphthalmia-associated transcription factor known as MITF (CACGTG, CATGTG), and E-boxes (CANNTG). Sex-determining region box 9 (or SOX9) response element previously reported in the regulation of RPE genes (including RPE65) was also identified in the newt RPE65 promoter. Third, we identified DNA motif boxes in the newt RPE65 promoter that are conserved among other vertebrates. The newt RPE65 promoter is an invaluable tool for site-specific delivery of exogenous genes or genetic manipulation systems for the study of retinal regeneration in this animal. |
| Starting Page | 463 |
| Ending Page | 473 |
| Page Count | 11 |
| File Format | |
| ISSN | 09628819 |
| Journal | Transgenic Research |
| Volume Number | 24 |
| Issue Number | 3 |
| e-ISSN | 15739368 |
| Language | English |
| Publisher | Springer International Publishing |
| Publisher Date | 2014-12-10 |
| Publisher Place | Cham |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Newt RPE65 Retinal pigment epithelium Transgenesis Molecular Medicine Plant Genetics & Genomics Animal Genetics and Genomics Plant Sciences Transgenics |
| Content Type | Text |
| Resource Type | Article |
| Subject | Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science |
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