Isolation and mapping of novel candidate genes for retinal disorders usingsuppression subtractive hybridization

We have constructed human cDNA libraries enriched for retina- and retinal p igment epithelium (RPE)/choroid-specific cDNAs through suppression subtract ive hybridization. The sequence of 314 cDNAs from the retina enriched libra ry and 126 cDNAs from the RPE/choroid enriched library was analyzed. Based on the absence of a database match, 25% of the retina cDNA clones and 16% o f the RPE/choroid cDNA clones are novel cDNAs. The expression profiles of 8 6 retina and 21 RPE/choroid cDNAs were determined by a semiquantitative rev erse transcription polymerase chain reaction technique. Thirty-three cDNAs were expressed exclusively or most prominently in retina or RPE/choroid. Th ese cDNAs were mapped in the human genome by radiation hybrid mapping. Elev en cDNAs colocalized with loci involved in retinal disorders. One cDNA mapp ed in a 1.5-megabase critical region for autosomal recessive retinitis pigm entosa (RP12). Another cDNA was assigned to the 7.7-cM RP17 linkage interva l. Seven cDNAs colocalized with four loci involved in Bardet-Biedl syndrome . (C) 1999 Academic Press.