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turnover of mRNA:gene transfection with new physical equipment (microporation)

November 30, 2009 - 07:25 -- Usa Lek-Uthai

Transfection system which is a new tool for RNAi (gene silencing) workflow; the study of human genome of 30000-40000 encoded protein , the protein function(s) are unknown or a little is known. the former era was studied on molecular biology which is the mutant phenotype sudies (Forward genetic). Recently there is reverse genetic which will rapidly results and clarify such as;

1. Gene over expression (phenotype level study)
2. Dominant negative expression (the protein malfunction study)
3. Gene knockout (the deletion mutation at the study gene on chromosome of live cell (animal model or cell lines) gene knockout for the study of drug development (enzymatic activity) this method cost very expensive and they may cause the death of growth cell or since embryonic stage).
4. antisense or ribosome (the study of turn over of mRNA (recommend)

The study of gene function using mRNA (Gene silencing or transcriptional gene silencing (PTGS) which is the transfection of transgene or double-stranded RNA(dsRNA) to the host cell and introduce the suppression of gene expression of target gene are hot issues. RNAi (cellular defense mechanism or innate cellular immune system ) which will destruct the target mRNA, will break the translation process, refer of gene malfunction. However, the only antisense siRNA use will complement to mRNA of target gene in host cell, then mRNA target will be destroyed by the nuclease enzymatic activity which is the RISC complex component. The transfection technique have many working flow, but the stealth RNA (chemical synthesis or siRNA (specific RNA Oligo 25bp-design by using software) which can reduce the stress of nonspecific response in cell, and how could we transfect this interesting RNA Oligo to cell, this can use an appropriate electric equipment (electrolytic buffer and pulse conditions [Pulse voltage/Pulse width/Pulse no.]). Such introduction of steath RNAi actin to cell, this stealth will depress the beta-actin synthesis of beta-actin gene (access by Realtime PCR). So transfection system is very easy and high transfection efficiency reported. Please see the transfection system on web. (many brands to compare).