Generation of kidney transcriptomes using serial analysis of gene expression.
Chronic renal disease initiation and progression remain incompletely understood. Genomewide expression monitoring should clarify the mechanisms which cause progressive renal disease by determining how clusters of genes coordinately change their activity. Serial analysis of gene expression (SAGE) is a technique of expression profiling which permits simultaneous and quantitative analysis of 9- to 13-bp sequence tags that correspond to unique mRNAs. Key principles of the technique are use of PCR in a manner to minimize distortion and serial concatenation of tags which facilitates sequencing and permits identification of many expressed genes in a single cDNA molecule. Tags are extracted from many concatenated sequences, counted using software, and identified by comparison with existing gene databases. In aggregate, gene expression profiles generated from a tag library comprise a transcriptome which represents a comprehensive and quantitative profile of genes expressed at the time of analysis. These global snapshots of gene expression patterns can better define basic cell biology and provide insights into disease pathogenesis by simultaneously determining the net consequences of gene-gene and gene-environment interactions on expression of thousands of genes. Rather than applying a priori assumptions (i.e., hypothesis testing), transcriptome analysis is hypothesis generating and requires no prior knowledge of gene expression. SAGE kidney transcriptomes, from normal animals and animals with progressive kidney disease, are being produced and can be analyzed for novel pathogenetic mechanisms. The use of SAGE and other genomic and proteomic tools should result in a better understanding of kidney disease pathogenesis and in identification of new therapeutic targets.
Schelling
Jeffrey R
JR
Department of Medicine, Rammelkamp Center for Education and Research, Case Western Reserve University, MetroHealth Medical Center Campus, 2500 MetroHealth Drive, G531, Cleveland, OH 44109-1998, USA.
El-Meanawy
M Ashraf
MA
Barathan
Shrinath
S
Dodig
Tatiana
T
Iyengar
Sudha K
SK
Sedor
John R
JR
D000818:Animals; D020869:Gene Expression Profiling; D023281:Genomics; D006801:Humans; D007668:Kidney / Q000502:physiology*; D014158:Transcription, Genetic / Q000235:genetics*