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B. Zangerl, J. Pillardy, B. Miller, Q. Sun, J. Johnson, P.A. Schweitzer, A.G. Hernandez, L. Liu, G.M. Acland, G.D. Aguirre; Development of a Normalized Canine Retinal cDNA Library for Microarray Analysis and Characterization of 10,000 ESTs . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2256.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Progressive Retinal Atrophy (PRA) in dogs, and Retinitis Pigmentosa (RP) in humans, comprises a clinical grouping of blinding hereditary diseases with broadly similar features. Despite the evidence that photoreceptor cells die via a common apoptotic pathway, little is known about the molecular mechanisms that trigger degenerative processes in different subsets of these disorders. To examine the cellular pathways related to retinal development and degeneration, we are currently establishing an integrated database on canine retinal expressed ESTs. Methods: We created and sequenced a canine normalized retinal cDNA library according to standard protocols using a modified phredPhrap algorithm for data analysis. Results: 9,984 cDNA clones yielded 6,557 high quality sequences which made up about 4,300 mostly non-redundant expressed ESTs, and showed an expected redundancy rate of approximately 40%. Approximately 4,000 non-redundant elements were identified for printing microarray chips as a tool to compare global gene expression between the normal and different non-allelic mutant retinas. Each represented element will be further characterized by homology search to other species and/or genomic location based on RH mapping. Conclusions: We created a normalized canine retinal cDNA library for investigation of expression patterns in normal and diseased retinas. To further sequence the library and expand the search for unique sequences for expanded microarray studies, the already characterized clones from the normalized retinal cDNA library will be subtracted to reduce the redundancy. Data obtained through different parts of this project are entered into an integrated database, an essential first step to establish an integrated network for gene identification and expression patterns suitable for comparative genetics, evolutionary analysis of gene families and functional genomics with respect to the developmental and degenerative processes of the retina.
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