Real-time gene expression analysis

We may also find that Sabatini s reverse transfection method of creating "live cell" microarrays offers even greater advantage for drug discovery and development (Ziauddin and Sabatini, 2001). Their method, relying on arrayed cDNA expression vectors to transform adherent cells, provides localized real-time gene expression analysis of the putative gene product. 

POTENTIAL APPLICATIONS FOR THE USE OF LANTHANIDE COMPLEXES 413 4. Real-time Gene Expression Analysis... 

Adjunctive to flux control analysis, other components of metabolism that contribute to product accumulation are needed including (1) substrate/precursor pool sizes (metabolomics), (2) co-factor capacities (metabolomics), (3) gene expression profiles (transcriptomics and quanfifafive real-time PCR), (4) protein profiles (pro-... 

O.A. Carmi, A.B. Stewart, S. Ulitzer, and J. Kuhn. 1987. Use of bacterial ludferase to establish a promoter probe vehicle capable of nondestructive real-time analysis of gene expression in Bacillus spp. i, Bacteriol. 169 2165-2170. 

Raman spectroscopy can offer a number of advantages over traditional cell or tissue analysis techniques used in the field of TE (Table 18.1). Commonly used analytical techniques in TE include the determination of a specific enzyme activity (e.g. lactate dehydrogenase, alkaline phosphatase), the expression of genes (e.g. real-time reverse transcriptase polymerase chain reaction) or proteins (e.g. immunohistochemistry, immunocytochemistry, flow cytometry) relevant to cell behaviour and tissue formation. These techniques require invasive processing steps (enzyme treatment, chemical fixation and/or the use of colorimetric or fluorescent labels) which consequently render these techniques unsuitable for studying live cell culture systems in vitro. Raman spectroscopy can, however, be performed directly on cells/tissue constructs without labels, contrast agents or other sample preparation techniques. 

Reverse transcription-polymerase chain reaction (RT-PCR) has been widely used for the detection of cytokine gene expression in clinical samples (W18, K5). However, conventional RT-PCR only offers a semiquantitative analysis. Recently, the Perkin-Elmer Corporation (Wellesley, MA) developed the TaqMan cytokine gene expression plate for real-time, in vitro quantitative evaluation of a panel of human cytokine gene expression using fluorescence detection. 

Chang TK, Chen J, Pillay V, Ho JY, Bandiera SM. Real-time polymerase chain reaction analysis of CYP1B1 gene expression in human liver. Toxicol Sci 2003 71 11-19. 

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25 402-408... 

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