Our Custom c-fos Serum Inducible Promoter System based Evaluation streamlines preclinical processes via advanced transcriptional pulsing technology. Leveraging the immediate early gene c-fos's natural, rapid, transient induction enables synchronized transcription initiation and a precise chase phase to measure true mRNA decay kinetics. We deliver definitive, physiologically relevant data beyond simple methods, optimizing mRNA sequence architecture to reduce immunogenicity, control therapeutic windows, and de-risk your pipeline with regulatory-ready insights.
The c-fos Serum Inducible Promoter System works by utilizing the inherent property of the c-fos immediate early gene, whose promoter is rapidly and transiently activated by serum stimulation to initiate synchronized transcription of the target mRNA. After this brief induction phase, transcription is naturally shut off, entering a "chase" period where no new mRNA is synthesized. By measuring the remaining abundance of the target mRNA at predefined time points during this chase phase, the system quantifies its natural decay rate and calculates the half-life, providing a physiologically relevant readout of mRNA stability without relying on transcription inhibitors.
Fig.1 Extracellular stimuli and calcium influx activate signaling pathways (e.g., MAPK, CaMK-CREB), triggering c-fos gene transcription via promoter-binding proteins; the resulting c-fos protein forms the AP-1 heterodimer with c-Jun and binds target gene DNA to exert effects.1
The primary detection indicators measured by our system are designed to offer a complete kinetic and mechanistic profile of mRNA turnover:
We employ a combination of gold-standard and next-generation analytical techniques to ensure maximal accuracy and sensitivity:
Our process is designed to be highly consultative and transparent, guiding your project from sequence submission to final, actionable stability metrics.
Client provides full mRNA sequence construct (including UTRs and coding sequence), purified plasmid DNA with expression cassette, and specific target cell line details (e.g., type, host species).
Create a chimeric reporter gene construct by cloning test destabilizing elements under the c-fos promoter. Prepare, validate, and serum-starve cells to achieve quiescent status.
Activate the c-fos promoter rapidly and transiently via serum induction to initiate synchronous transcription of target mRNA. Collect RNA samples at precise pre-determined time points (the "chase" phase).
Isolate total RNA and analyze using high-sensitivity detection methods. Use raw RNA data to calculate the half-life (t1/2) and decay kinetics of the novel transcript.
Compile raw data, calculate half-life values, comparative analysis against controls, and deliver a comprehensive report.
The typical timeframe for this comprehensive evaluation ranges from 5 to 8 weeks, depending on the complexity of the mRNA construct and the need for upstream vector engineering.
Discover How We Can Help - Request a Consultation
As a trusted partner in advanced therapeutic development, Creative Biolabs is dedicated to providing bespoke solutions that meet the rigorous demands of your preclinical pipeline. Our service for c-fos Serum inducible Promoter System based Evaluation is an advanced, high-fidelity offering designed for the modern biology expert.
Physiological Fidelity
We use the natural, transient c-fos promoter response, eliminating the artifacts associated with transcription inhibitors and guaranteeing stability data that mimics true in vivo behavior.
Unparalleled Synchronization
The rapid transcriptional pulse ensures that mRNA synthesis is repressed with a high degree of synchrony, allowing for the precise and unambiguous measurement of decay kinetics for both unstable and long-lived transcripts.
Customized Contextual Testing
We offer the flexibility to run the evaluation in client-specified mammalian cell lines (provided they are c-fos responsive), guaranteeing the data is relevant to your specific therapeutic delivery and target context.
One-Stop Mechanistic Evaluation
Our integrated platform covers the entire process, from chimeric construct design to the final quantitative half-life report, including specialized Northern Blotting for unequivocal deadenylation status determination.
Regulatory-Ready Deliverables
Our well-established quality system and commitment to high-standard QC tools ensure the generated stability data is quantitative, reliable, and suitable for supporting your IND-enabling studies and regulatory submissions.
Experience the Creative Biolabs Advantage - Get a Quote Today
Actinomycin D stops all cellular transcription, which can introduce non-physiological stress and potentially toxic artifacts that skew the stability data. Our c-fos system, on the other hand, utilizes a natural, transient, serum-based transcriptional pulse. This provides a clean, highly synchronized starting point for the decay chase, yielding more accurate and physiologically relevant half-life measurements. This is a major advantage for de-risking your therapy.
Yes, absolutely. The highly synchronous nature of the c-fos transcriptional pulse is designed to accurately measure both extremes. The sharp initial pulse makes it easier to capture the rapid decay kinetics of unstable transcripts, while the sustained duration of the chase phase allows for the precise measurement of even relatively long-lived mRNAs.
We specialize in customization. While the system is highly effective in standard lines, we can work with many client-provided cell lines, provided they exhibit the necessary c-fos responsiveness. We perform a preliminary validation to ensure the system's tight regulatory control is maintained, allowing you to test your therapeutic in a context most relevant to your final clinical application.
Our service is designed to evaluate the intrinsic stability of the mRNA molecule itself. While LNP formulation is a separate service, determining the optimal inherent sequence stability first with our c-fos system is the foundational step that ensures the LNP is encapsulating the highest-quality, most stable transcript possible. We recommend starting here before full-scale LNP production.
Creative Biolabs' c-fos Serum inducible Promoter System based Evaluation is the strategic choice for drug developers who cannot afford ambiguity. By integrating molecular biology expertise with a cutting-edge transcriptional pulsing platform, we deliver definitive, quantitative mRNA stability data that accelerates therapeutic development, informs critical molecular architecture decisions, and supports robust regulatory submissions.
Contact Our Team for More Information and to Discuss Your Project
Reference
