Isolation and mapping of novel candidate genes for retinal disorders using suppression subtractive hybridization.
We have constructed human cDNA libraries enriched for retina- and retinal pigment epithelium (RPE)/choroid-specific cDNAs through suppression subtractive hybridization. The sequence of 314 cDNAs from the retina enriched library 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% of the RPE/choroid cDNA clones are novel cDNAs. The expression profiles of 86 retina and 21 RPE/choroid cDNAs were determined by a semiquantitative reverse transcription polymerase chain reaction technique. Thirty-three cDNAs were expressed exclusively or most prominently in retina or RPE/choroid. These cDNAs were mapped in the human genome by radiation hybrid mapping. Eleven cDNAs colocalized with loci involved in retinal disorders. One cDNA mapped in a 1.5-megabase critical region for autosomal recessive retinitis pigmentosa (RP12). Another cDNA was assigned to the 7.7-cM RP17 linkage interval. Seven cDNAs colocalized with four loci involved in Bardet-Biedl syndrome.
den Hollander
A I
AI
Department of Human Genetics, University Hospital Nijmegen, Nijmegen, 6500 HB, The Netherlands. A.denHollander@antrg.azn.nl
van Driel
M A
MA
de Kok
Y J
YJ
van de Pol
D J
DJ
Hoyng
C B
CB
Brunner
H G
HG
Deutman
A F
AF
Cremers
F P
FP
D000818:Animals; D002829:Choroid / Q000378:metabolism; D002874:Chromosome Mapping; D006224:Cricetinae; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D005786:Gene Expression Regulation; D015723:Gene Library; D005796:Genes / Q000235:genetics; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D010857:Pigment Epithelium of Eye / Q000378:metabolism; D012164:Retinal Diseases / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary