With decades of experience in mRNA research, Creative Biolabs became a trusted partner to provide mRNA services. Our scientists have developed a variety of different systems for mRNA stability tests to meet your objectives, such as the c-fos serum-inducible promoter system.
Among the genes most studied concerning, mRNA stability is the protooncogenes involved in the early response to growth factors, especially c-fos. Transcription of c-fos is rapidly induced by growth factors and other extracellular stimuli and then ceases completely within minutes after its activation. This transient accumulation and tight control of c-fos mRNA appears to be critical to normal cellular function. Moreover, monitoring this rapid decay has permitted the direct measurement of c-fos mRNA turnover under conditions of serum stimulation. This approach provides a tight genetic switch to control transcription for studying mRNA turnover without the use of transcription inhibitors.
Along these years, there are many different methods developed for measuring mRNA stability, for example, those transcriptional pulsing methods based on the c-fos serum-inducible promoter can better elucidate the mechanistic steps and regulation underlying differential and selective mRNA turnover in mammalian cells. Creative Biolabs provides c-fos serum-inducible promoter system that allows unequivocal monitoring of deadenylation and decay kinetics as well as the determination of precursor-product relationship to denote the mRNA stability quantitatively. Thus, the c-fos serum-inducible promoter system provides a reliable way to test physiologically meaningful changes in mRNA stability.
The method takes advantage of the quick and transient response of the c-fos promoter to serum induction in mammalian cells. Usually, a chimeric reporter gene is first constructed in which mRNA destabilizing elements can be introduced and subsequently tested. After individually introducing test destabilizing sequences into the plasmid construct, the construct is delivered to cells by transient transfection assay. Cells are subsequently serum-starved to make them quiescent. The c-fos promoter is capable of responding transiently to a shift from low to high serum-containing medium. Thus, gene transcription driven by the c-fos promoter is transiently induced by the addition of serum. mRNA decay and the half-life can be determined simply by monitoring the subsequent rate of mRNA loss. To do this, RNA samples are collected at different time points after serum induction to determine the decay rate of the mRNAs by Northern blot analysis. This system is efficient and allows the analysis of transcripts that derive from transiently introduced genes so that it eliminates the need to establish stably transfected cell clones.
The use of a regulatable c-fos serum-inducible promoter represents a major improvement in the analysis of eukaryotic mRNA decay and mRNA stability. Creative Biolabs provides the c-fos serum-inducible promoter system to determine mRNA stability for our customers' specific requirements. If you are interested in our services, please do not hesitate to contact us or directly send us an online inquiry.