mRNA Stability Analysis Using Actinomycin D Assay
mRNA stability analysis within cells provides important information in understanding post-transcriptional gene regulation as well as the potential role of RNA-protein interactions. To measure mRNA stability within cells, one of the simplest techniques is to inhibit transcription in vivo with transcription inhibitors and then measure the mRNA kinetics. Actinomycin D is a transcription inhibitor and widely used in mRNA stability within mammalian cells. The assay allows the assessment of mRNA decay by determining mRNA abundance following transcription inhibition. With years of experience in mRNA stability assay, Creative Biogene is proud to offer mRNA stability analysis services using actinomycin D assays to satisfy our global customers' specific project needs. Our services include measuring and analysis of mRNA stability for cultured mammalian cells, insect cells, as well as exogenously-expressed transcripts. Aided by our mRNA stability platforms and experienced experts, researchers can rapidly and extensively investigate mRNA stability to understand gene expression changes as well as the underlying mechanisms that regulate the level of transcripts at different physiological conditions.
Actinomycin D assay
Actinomycin D assay as a transcription inhibition method has probably the longest history of determining mRNA stability under various experimental conditions in cultured cells. This assay has the capability to measure stability changes of endogenous mRNAs without the requirement of the construction and introduction of exogenous genes into cells. Actinomycin D is an antibiotic derived from Streptomyces parvullus. As a transcription inhibitor, the action mechanism of Actinomycin D is complex, but the major mechanism is believed to involve direct intercalation of DNA, following inhibition of RNA polymerases (RNAP) and transcription decrease. Although the actinomycin D method has several disadvantages, such as being prone to artifacts and ineffective for some promoters. The method can be readily applied on a genome-wide level and is highly cost-effective. We believe that this technique for measuring endogenous mRNA stability can serve as a versatile, effective tool under appropriate guidelines and controls.
mRNA stability analysis using actinomycin D assay
With appropriate guidelines and controls, and an actinomycin D-based method is a powerful tool for measuring mRNA decay rates. Here, we offer a one-stop solution for mRNA stability analysis using actinomycin D assay. Notably, actinomycin D is toxic. We have a specialized laboratory and experienced technicians to store and handle it.
In general, our services include four steps, including cell cultures, cell treatment with actinomycin D, mRNA quantitation, and results analysis. Among them, cell treatment with actinomycin D is the most important step. Different cultured cells have differential sensitivities to the cytotoxic effects of actinomycin D. For new cell types of our customers, we usually perform a time course of cytotoxicity with several actinomycin D concentrations and the results will be considered for mRNA stability analysis. Moreover, the success of an actinomycin D experiment relies on the methods used to measure mRNA concentrations. We offer a series of methods for customers to choose from, including northern blot, real-time RT-PCR, and RNA-Seq. We will offer the optimal strategy according to customers' budgets and project requirements.
Features of our service include
- An optimized and well-established platform.
- Professional technical support.
- One-stop solutions.
- Ready to start your project once the contract is signed.
Your success is important to all of us, and our mRNA stability analysis service is supported by years of experience in the development of an optimized and well-established platform. Simply let us know about your research needs. We will propose the best strategy to best match your research objectives. Contact us right now!
- Wada, T., & Becskei, A. (2017). "Impact of Methods on the Measurement of mRNA Turnover." International journal of molecular sciences, 18(12), 2723.