English version
Instituts/ Institut de biologie et de technologies de Saclay (iBiTec-S)/ Unités/ Service de biologie intégrative et génétique moléculaire (SBIGeM )/ Laboratoire de physio génomique (LPG)/ Adaptation du SAGE pour micro-quantités

Adaptation du SAGE pour micro-quantités



Let's notice in Paris by the end of the XVIIIth century (SADE) and
by the end of the XIXth century (LAUTRÉAMONT) a brief
appearance of the philosophers' stone.
RENÉ CHAR (Tribute to D.-A.-F. de SADE)

If your are looking for detailed information about SADE or want to cite this work, please refer to these articles :

  1. SADE: a microassay for serial analysis of gene expression
    Cheval L., Virlon B., and Elalouf J.-M.  In: Functional Genomics: a pratical approach. Edited by S.Hunt and J.-P. Livesey. Oxford University Press 2000 pp. 139-163
     
  2. Serial microanalysis of renal transcriptomes
    Virlon B., Cheval L., Buhler J.-M., Billon E., Doucet A., and Elalouf J.-M. Proc. Natl. Acad. Sci. USA 96:15286-15291, 1999 

CEA/CNRS has licensed the SADE technology for commercial purposes, but it is freely available to academia for research purposes.



Serial microanalysis of renal transcriptomes

Berangere VIRLON, Lydie CHEVAL, Jean-Marie BUHLER, Emmanuelle BILLON, Alain DOUCE

and Jean-Marc ELALOUF


 Departement de Biologie Cellulaire et Moleculaire, CNRS URA 1859, CEA Saclay, 91191 Gif-sur-Yvette Cedex, France

Large scale gene expression studies can now be routinely performed on macroamounts of cells, but it is unclear to which extent current methods are valuable for analyzing complex tissues. In the present study, we used the method of Serial Analysis of Gene Expression (SAGE) for quantitative mRNA profiling in the mouse kidney. We first performed SAGE at the whole kidney level by sequencing 12,000 mRNA tags. Most abundant tags corresponded to transcripts widely distributed or enriched in the predominant kidney epithelial cells (proximal tubular cells), whereas transcripts specific for minor cell types were barely evidenced. To better explore such cells, we set up a SAGE Adaptation for Downsized Extracts (SADE), enabling a 1,000-fold reduction of the amount of starting material. The potential of this approach was evaluated by studying gene expression in microdissected kidney tubules (50,000 cells). Specific gene expression profiles were obtained, and known markers (e.g. uromodulin in the thick ascending limb of Henle's loop, and aquaporin-2 in the collecting duct) were found appropriately enriched. In addition, several enriched tags had no databank match, suggesting that they correspond to unknown or poorly characterized transcripts with specific tissue distribution. It is concluded that SADE makes possible large scale quantitative gene expression measurements in small biological samples, and will help to study the tissue expression and function of genes not evidenced with other high-throughput methods.




SADE data

  1. Serial microanalysis of renal transcriptomes
    Virlon B, Cheval L, Buhler JM, Billon E, Doucet A and Elalouf JM
    Proceedings of the National Academy of Sciences USA, 1999, 96: 15286-15291

  2. Gene expression profiles in normal and Otx2-/- early gastrulating mouse embryos
    Zakin L, Reversade B, Virlon B, Rusniok C, Glaser P, Elalouf JM and Brûlet P
    Proceedings of the National Academy of Sciences USA, 2000, 97: 14388-14393
  3. Transcriptome of a mouse kidney cortical collecting duct cell line: effects of aldosterone and vasopressin
    Robert-Nicoud M, Flahaut M, Elalouf JM, Nicod M, Salinas M, Bens M, Doucet A, Wincker P, Artiguenave F, Horisberger JD, Vandewalle A, Rossier BC and Firsov D
    Proceedings of the National Academy of Sciences USA, 2001, 98(5): 2712-2716



JME, JCA