Identification of candidate genes for drug discovery by differential display

✍ Scribed by Lily Shiue

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 142 KB
• Volume: 41
• Category: Article
• ISSN: 0272-4391
• DOI: 10.1002/(sici)1098-2299(199707/08)41:3/4<142::aid-ddr5>3.0.co;2-l

✦ Synopsis

Regulation of gene expression can specify cellular fate, define responses to stimuli, and contribute to complex microenvironments present in tissues. Identification of differentially expressed genes in experimental paradigms can help elucidate underlying biochemical pathways and thus reveal potential therapeutic targets. The technique of differential display uses arbitrarily primed PCR to sample complex cDNA populations of interest; amplified portions of messenger RNAs are analyzed by denaturing gel electrophoresis and those which are differentially represented can be directly visualized and cloned. PCR-based techniques for analysis of gene expression are reliable and extremely sensitive. In comparison to traditional methods, such as subtractive hybridization, differential display allows for many samples to be compared in parallel, and the requirement for starting material is low. There are a plethora of examples in the literature of how differentially expressed genes can be rapidly identified in experimental paradigms ranging from cells treated in culture to whole organs of treated animals. The challenge for the researcher is then defining candidate genes for drug discovery from an initial screen based only on differential expression patterns. Careful experimental design and execution are critical for optimal use of such methodologies to fill a gene discovery pipeline. In this article, the merits and potential pitfalls of differential display and related PCRbased techniques are discussed. Current protocols are reviewed and innovations pertaining to high-throughput applications are noted. Drug Dev. Res. 41:142-159, 1997.