Retinal development, function, and disease are, to a significant degree, controlled by the pattern of genes expressed by the cells of the retina. In an effort to better understand the mechanisms regulating photoreceptor gene expression, we have been studying rod and cone opsin gene regulation as model systems. Using a variety of approaches, we, and others, have defined some of the DNA elements important for rod- and cone-specific expression, have identified and cloned some of the transcription factors that bind to these DNA elements, and have shown that mutations in some of these factors can both interfere with normal photoreceptor development in the mouse and can cause retinal degeneration in man. This application for continued funding of these studies proposes to continue and broaden this work. The proposed work includes four aims: 1) more fully characterize several already cloned factors that are candidate molecules for regulating the various cone opsins;2) further develop and utilize in vitro retinal electroporation technology so as to assess the biological activity of rod and cone opsin regulatory elements in a more physiological milieu, and in a complementary fashion use a high-throughput PCR-based DNase I hypersensitivity assay to identify regions of regulatory interest;3) utilize a high- throughput transient transfection-based assay together with libraries of transcription factor expression clones to identify factors that regulate the response elements identified in "2";and 4) use a combined bioinformatic and experimental approach to identify genes that are differentially expressed in the retina, and then determine how expression of these retina-specific splice isoforms varies during development, during degeneration in the rd1 mouse, and in three mouse models that express altered forms of three pre-RNA processing factors (PRPF 3, 8 and 31) that when altered in humans cause retinitis pigmentosa. Together, these studies will hopefully provide new insights into the mechanisms regulating photoreceptor gene expression, which in turn will aid in understanding how abnormalities in gene expression can cause retinal disease, and help in the development of more cell-type specific gene therapy approaches.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY009769-20
Application #
8204531
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Neuhold, Lisa
Project Start
1992-08-01
Project End
2013-11-30
Budget Start
2012-03-01
Budget End
2013-11-30
Support Year
20
Fiscal Year
2012
Total Cost
$580,672
Indirect Cost
$226,604
Name
Johns Hopkins University
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Elachouri, G; Lee-Rivera, I; Clérin, E et al. (2015) Thioredoxin rod-derived cone viability factor protects against photooxidative retinal damage. Free Radic Biol Med 81:22-9
Wan, Jun; Oliver, Verity F; Wang, Guohua et al. (2015) Characterization of tissue-specific differential DNA methylation suggests distinct modes of positive and negative gene expression regulation. BMC Genomics 16:49
Masuda, Tomohiro; Berlinicke, Cynthia A; Zack, Donald J (2015) Gene regulation: it matters who you hang out with. Neuron 86:7-9
Hiebler, Shandi; Masuda, Tomohiro; Hacia, Joseph G et al. (2014) The Pex1-G844D mouse: a model for mild human Zellweger spectrum disorder. Mol Genet Metab 111:522-32
Masuda, Tomohiro; Wahlin, Karl; Wan, Jun et al. (2014) Transcription factor SOX9 plays a key role in the regulation of visual cycle gene expression in the retinal pigment epithelium. J Biol Chem 289:12908-21
Wahlin, Karl J; Maruotti, Julien; Zack, Donald J (2014) Modeling retinal dystrophies using patient-derived induced pluripotent stem cells. Adv Exp Med Biol 801:157-64
Masuda, Tomohiro; Zhang, Xiaodong; Berlinicke, Cindy et al. (2014) The transcription factor GTF2IRD1 regulates the topology and function of photoreceptors by modulating photoreceptor gene expression across the retina. J Neurosci 34:15356-68
Ranganathan, Vinod; Wahlin, Karl; Maruotti, Julien et al. (2014) Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide RNAs. Nat Commun 5:4516
Sluch, Valentin M; Zack, Donald J (2014) Stem cells, retinal ganglion cells and glaucoma. Dev Ophthalmol 53:111-21
Bhise, Nupura S; Wahlin, Karl J; Zack, Donald J et al. (2013) Evaluating the potential of poly(beta-amino ester) nanoparticles for reprogramming human fibroblasts to become induced pluripotent stem cells. Int J Nanomedicine 8:4641-58

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