mRNA's 5' cap (m7GpppG) aids stability/translation, but standard IVT mRNA is prone to decapping and degradation. The 6-Thioguanosine (s6G) Cap, a modified analog, resists decapping enzymes and boosts eIF4E recruitment, enhancing mRNA half-life and efficacy (backed by literature).
Creative Biolabs' Custom s6G Cap Service uses next-gen s6G analogs to solve long development cycles, low yields, and instability. It boosts mRNA stability/translation, increases therapeutic protein production, and maximizes mRNA constructs' therapeutic potential to accelerate programs.
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Fig.1 The molecular structure of 6-thioguanine (6-TG).1
The 6-Thioguanosine (s6G) cap is a derivative of guanosine where the oxygen atom at the C6 position of the guanine base is replaced by a sulfur atom. This modification is incorporated at the 5' terminus via a triphosphate linkage (e.g., s6GpppG). The resulting structure is similar enough to the native cap to bind eIF4E effectively, but the substitution confers critical functional differences.
The s6G cap's mechanism of action is two-fold:
The s6G cap provides key advantages for therapeutic development:
The 6-Thioguanosine-containing cap is widely utilized in projects requiring maximum stability and quantitative data:
The delivery of a high-purity, next-generation mRNA construct requires a robust, validated workflow suitable for visualization as a standard operating procedure (SOP).
| Starting Materials | Key Steps Involved |
|---|---|
| mRNA Template: Linearized DNA plasmid or specific IVT template sequence. | Project Consultation & Design: Comprehensive review of the client's therapeutic goals, scale requirements, and final application. Selection of the optimal capping method (co-transcriptional vs. enzymatic post-transcriptional). |
| Scale and Purity: Required final mRNA scale (e.g., 10 mg or gram scale) and purity specifications. | Custom s6G Synthesis: In vitro transcription (IVT) utilizing high-fidelity polymerases and customized s6G cap analogs to generate the desired mRNA construct. |
| Application Context: Target cell type (e.g., T-cells, Dendritic Cells) and planned in vivo model. | Purification & Quality Control (QC): Multi-step purification (e.g., HPLC) to remove truncated transcripts, enzymes, and unincorporated nucleotides. QC includes integrity analysis and concentration validation. |
| Cap Verification & Functionality Check: Mass Spectrometry (MS) and HPLC are used to verify the correct molecular weight and s6G incorporation efficiency. Stability assays may be run to confirm Dcp1/2 resistance. | |
| Final Formulation and Delivery: Final formulation optimized for stability (e.g., lyophilization or solution) and shipped with a comprehensive data package. |
Final Deliverables:
Estimated Timeframe: The typical timeframe for this custom service ranges from 4 to 8 weeks, depending critically on the required synthesis scale and the complexity of the custom quality control (QC) assays requested.
Leverage Creative Biolabs' expertise to transform your therapeutic mRNA with our high-impact 6-Thioguanosine-containing Cap Service. We provide elite, flexible solutions tailored precisely to your project's phase and scale.
Customized Production Scale
We offer a seamless transition from laboratory-scale synthesis (milligrams for early research) up to gram-scale production batches required for preclinical and early-phase clinical studies.
Elite Cap Efficiency
Guaranteed high-efficiency capping (approaching 100%) achieved through optimized in vitro transcription (IVT) processes that minimize the formation of inactive reverse-cap structures.
End-to-End Process Control
Efficient upstream development (custom IVT) and downstream purification (HPLC) ensure the final product is a single, highly pure RNA species suitable for demanding delivery systems like LNPs.
Advanced Quality-by-Design (QbD)
We implement rigorous quality controls, including Mass Spectrometry (MS) and HPLC, to accurately quantify and evaluate the quality of the modified mRNA, ensuring data integrity for regulatory readiness.
Custom s6G Solutions
Our service is customized to your specific mRNA sequence and target application, allowing optimization of codon usage and 5'/3' features to facilitate maximum expression in selected cell types.
Analytical Support
Access to the unique photo-crosslinking capabilities of s6G, providing a superior tool for structural analysis and cap-protein interaction studies critical for mechanism-of-action validation.
A: Not at all. The s6G cap is a precision molecular modification designed to maintain native function while enhancing stability. The cap itself is sequestered within the Lipid Nanoparticle (LNP) until cellular uptake, ensuring it does not negatively impact particle formation, surface chemistry, or targeting. The only difference you'll see is a more stable, functional payload once it's released in the cell.
A: ARCA is an excellent tool for ensuring correct 5' orientation, but the s6G cap offers a superior level of protection against cellular degradation because it is chemically resistant to the Dcp1/2 decapping enzyme—a benefit ARCA alone does not provide. If your primary pain point is in vivo half-life and sustained expression, s6G is the clear next-generation choice.
A: Absolutely. In fact, s6G is highly advantageous for cell reprogramming. The increased stability and higher protein translation efficiency are critical when you need to drive robust, rapid changes in cell fate or function with minimal exogenous material. This approach minimizes cellular stress while maximizing therapeutic impact.
A: Yes, comprehensive MS verification is a standard, essential component of our QC package. We employ high-resolution mass spectrometry to confirm the correct molecular weight and structure of the s6G cap, providing you with unambiguous, defensible data that proves the integrity and quality of your final construct.
A: We are equipped to synthesize high-efficiency s6G-capped mRNA constructs up to gram scales, ensuring seamless continuity as you progress from bench-scale testing to large-animal studies and Phase I clinical batch requirements. The feasibility depends on the sequence length and complexity, which is precisely why we encourage a personalized consultation.
The key to unlocking the full potential of your mRNA therapeutic is precision engineering—both at the molecular level (with the s6G cap) and the delivery level. Our team at Creative Biolabs is ready to provide the custom solutions that will differentiate your product in the rapidly evolving market.
Contact Our Team for More Information and to Discuss Your Project| Cat. No | Product Name | Promoter |
|---|---|---|
| CAT#: GTVCR-WQ001MR | IVTScrip™ pT7-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ002MR | IVTScrip™ pT7-VEE-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ003MR | IVTScrip™ pT7-VEE-mRNA-FLuc Vector | T7 |
| CAT#: GTVCR-WQ87MR | IVTScrip™ pT7-VEE-mRNA-Anti-SELP, 42-89-glycoprotein Vector | T7 |
| Cat. No | Product Name | Type |
|---|---|---|
| CAT#: GTTS-WQ001MR) | IVTScrip™ mRNA-EGFP (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| CAT#: GTTS-WK18036MR | IVTScrip™ mRNA-Human AIMP2, (Cap 1, Pseudo-UTP, 120 nt-poly(A)) | Enzyme mRNA |
| (CAT#: GTTS-WQ004MR) | IVTScrip™ mRNA-Fluc (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| (CAT#: GTTS-WQ009MR) | IVTScrip™ mRNA-β gal (Cap 1, 30 nt-poly(A)) | Reporter Gene |
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