Genome-wide analysis of antisense transcription with Affymetrix exon array
1 Department of Mathematics and Statistics, South Dakota State University, Box 2220, Brookings, SD 57007, USA
2 Center for Medical Genetics, Evanston Northwestern Healthcare, 1000 Central Street, Suite 620, Evanston, IL 60201, USA
3 Center for Functional Genomics, ENH Research Institute, 1001 University Place, Evanston, IL 60201, USA
4 Dept. of Plant Pathology & Microbiology, College of Natural and Agricultural Sciences, University of California Riverside, Riverside, CA 92521, USA
5 Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA
BMC Genomics 2008, 9:27 doi:10.1186/1471-2164-9-27Published: 22 January 2008
A large number of natural antisense transcripts have been identified in human and mouse genomes. Study of their potential functions clearly requires cost-efficient method for expression analysis.
Here we show that Affymetrix Exon arrays, which were designed to detect conventional transcripts in the sense orientation, can be used to monitor antisense expression across all exonic loci in mammalian genomes. Through modification of the cDNA synthesis protocol, we labeled single-strand cDNA in the reverse orientation as in the standard protocol, thus enabling the detection of antisense transcripts using the same array. Applying this technique to human Jurkat cells, we identified antisense transcription at 2,088 exonic loci of 1,516 UniGene clusters. Many of these antisense transcripts were not observed previously and some were validated by orientation-specific RT-PCR.
Our results suggest that with a modified protocol Affymetrix human, mouse and rat Exon arrays can be used as a routine method for genome-wide analysis of antisense transcription in these genomes.