Our Custom mRNA Stability Evaluation Services de-risk therapeutic candidates via rigorous, high-throughput mRNA half-life measurement assays. mRNA lifespan directly determines in vivo therapeutic protein production, with variability/degradation derailing R&D efforts.
Creative Biolabs delivers definitive quantitative data to validate construct design, optimize formulation, and meet regulatory requirements. Leveraging gold-standard methods, we provide foundational kinetic intelligence for predictable therapeutics, streamlining clinical pathways.
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mRNA degradation is a highly regulated process influenced by several complex, coupled factors. These include the length and sequence of the Untranslated Regions (UTRs), the presence of specific structural motifs like G-quadruplexes or i-motifs, the codon optimality of the coding sequence, and the binding affinity of various RNA-binding proteins (RBPs) and ribonucleases (RNases). Our assays are designed to test stability across changes in these specific factors.
The primary detection indicator is the mRNA Half-Life (t1/2), which is the time required for half of the initial mRNA molecules to degrade. Other indicators often include the decay rate constant and the overall relative abundance of the target transcript compared to a stable internal control or housekeeping gene.
This is one of the most common and high-throughput methods. It involves applying the transcription inhibitor Actinomycin D to cells, rapidly halting all de novo mRNA synthesis. Subsequent measurements of the target mRNA over time are assumed to reflect the kinetics of degradation alone. This method is highly amenable to high-throughput screening and measuring the stability of endogenous or transiently expressed transcripts.
This system utilizes the c-fos serum-inducible promoter to initiate a rapid, pulsatile wave of reporter gene transcription upon serum stimulation. Since the induction is rapid and transient, the subsequent degradation of the mRNA can be tracked precisely without the toxicity associated with blanket transcription inhibitors like Actinomycin D. It is an excellent tool for studying degradation kinetics in mammalian tissue culture cells.
Inducible regulatory systems enable conditional and reversible control of gene expression. In such systems, the transcription of target genes remains inactive (or active) under basal conditions, and can be rapidly switched "on" or "off" upon addition of a specific inducer (e.g., small molecules, light, or metabolites). This tightly regulated mode supports precise investigation of gene function, protein dynamics, or pathway responses, as it avoids.
Recognizing that no single off-the-shelf assay fits every therapeutic development pipeline, we offer Custom Stability Evaluation Assay Development. Leveraging our advanced techniques and over two decades of experience, we custom-design and validate novel assays to meet unique project demands, such as testing stability in complex cell-free systems, specific organoid models, or under non-standard environmental conditions.
This detailed, professional workflow is suitable for visualization as a clear flowchart, making the entire process transparent to potential clients.
Based on the client's specific mRNA sequence, host system, and regulatory goals, our experts select and optimize the appropriate evaluation method (e.g., Actinomycin D or inducible systems). This ensures the assay is sensitive and relevant.
The client's mRNA, often delivered via the specified vector or as an in vitro transcribed (IVT) product, is introduced into the defined cellular system under the agreed-upon conditions.
Transcription of the target gene is rapidly halted or 'pulsed' using a highly characterized method (e.g., Actinomycin D addition or inducible system deactivation) to establish a clear decay starting point.
Multiple time points are carefully sampled over the expected degradation curve. Total RNA is extracted from each time point using high-quality, standardized protocols to minimize RNase activity artifacts.
The target mRNA is quantified at each time point using highly specific and validated reverse transcription quantitative polymerase chain reaction (RT-qPCR) or droplet digital PCR (ddPCR) to determine absolute transcript levels.
The resulting kinetic data points are plotted, normalized, and fitted to a first-order decay model to accurately calculate the empirical mRNA half-life under the tested conditions.
Upon completion, clients receive the comprehensive data package required for decision-making and regulatory filing:
Creative Biolabs delivers more than just data—we provide predictive kinetic intelligence through our highly customizable and robust platform. Our services are specifically designed to inject certainty into your genetic engineering and therapeutic development programs.
We design and validate bespoke stability protocols (e.g., Actinomycin D, inducible system, c-fos) specifically tailored to your therapeutic sequence, delivery vehicle, and unique host cell system.
Utilize automated platforms for massively parallel kinetic decay measurements across multiple sequence variants or complex conditions, drastically reducing crucial screening time.
Deliver precise, quantitative mRNA half-life t1/2 data using highly sensitive RT-qPCR and ddPCR, providing the definitive metrics essential for all regulatory filings.
Generate critical, empirical data on how your LNP, polyplex, or other formulation protects the mRNA against nucleases, enabling data-driven optimization of the delivery strategy.
Our high-fidelity kinetic data serves as the gold standard empirical input used to train and validate advanced in silico sequence-to-function stability models.
Provide comprehensive, audit-ready reports that strictly meet the data rigor required for IND-enabling studies and subsequent regulatory submissions.
There are methods to design and construct the mRNA sequences to be detected on a large scale through biophysical modeling and machine learning. After amplification, the dynamic mRNA levels are measured using DNA-Seq and RNA-Seq. mRNA sequences are designed based on a combination of multiple factors that affect the attenuation rate of mRNA, including pH, binding affinity, secondary and tertiary structures of mRNA, and the translation rate of mRNA.
Fig.1 Large-scale parallel design and dynamic attenuation measurement.1
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While in silico tools offer initial guidance, they often lack the fidelity needed for clinical translation. Creative Biolabs' service provides the empirical, quantitative mRNA Half-Life data derived from real cellular environments, using gold-standard RT-qPCR/ddPCR. This eliminates prediction error and is the definitive metric required for regulatory confidence.
That's a great question about data integrity. We prioritize clean kinetics by carefully titrating inhibitors and offering alternative, less toxic systems like the inducible system or c-fos promoter methods. We always optimize the protocol to your specific cell line to ensure the observed decay is purely a function of mRNA degradation, not cellular stress.
Absolutely. Our platform is built for High-Throughput Kinetic Screening. We can efficiently measure the t1/2 for dozens of sequence variants (including UTRs, structural motifs, and RBP-binding sites) in a single run. This saves you significant time in the Design-Build-Test cycle and allows for true rational design.
Yes, our focus is on Regulatory-Grade Reporting. We utilize validated, standardized protocols (like the inducible system system for clean control) and provide comprehensive documentation, including detailed assay reports and confidence intervals. Our metrics directly support the demonstration of controlled gene expression, which is a critical regulatory requirement.
We specialize in Custom Stability Evaluation Assay Development. We routinely adapt our methods to non-standard environments, including organoids, complex co-culture systems, and specific in vivo mimics. This critical customization ensures the stability of data is highly relevant to your true clinical application.
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Creative Biolabs offers the definitive Custom mRNA Stability Evaluation Services to empower your next-generation therapeutic development. Our advanced, custom-designed assays provide the reliable kinetic data needed to transition your project from the bench to the clinic with confidence.
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