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S Zareparsi, M Buraczynska, AJ Mears, R Farjo, SP MacNee, B Hughes, A Swaroop; Towards an Expression Profile of Native Human Retinal Pigment Epithelium and Identification of RPE-Specific Genes . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2434.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To generate a profile of genes expressed in the native human retinal pigment epithelium and to examine their tissue specific expression pattern to identify novel RPE-specific genes. Methods: An unamplified cDNA library was constructed using RNA isolated from native human RPE sheets. The sequence from the 5'end was obtained for randomly selected clones; of these, more than 1600 expressed sequence tags (ESTs) were analyzed for similarity to sequences and gene clusters in public databases. cDNA microarray slides containing RPE-expressed genes from this library will be generated and hybridized against RNA from RPE, retina and kidney. Results: EST analysis revealed several known RPE-expressed genes and more than 500 genes that have been characterized previously but not known to be expressed in the RPE. Over 200 novel ESTs and putative proteins have been identified. An additional 344 sequences only matched human genomic sequence. Comparative analysis of hybridizations with RNA from RPE, retina, and kidney will be performed in order to identify genes that are specifically expressed in the RPE. Conclusion: The analysis of ESTs generated from the native human RPE library has revealed a set of novel ESTs, putative proteins and genomic hits, which may represent as yet unidentified RPE-specific genes. Many of these map to the chromosomal regions of previously localized disease loci and may serve as candidate genes. Identification of novel RPE-specific genes will lead to a better understanding of the role that RPE plays in normal eyes and in disease state. In particular, RPE-specific genes will be studied as candidates for susceptibility to age-related macular degeneration.
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