2-Thiouridine Modification Service

Introduction

Post-transcriptional RNA modifications are conserved for regulating gene expression. 2-Thiouridine (s²U), a natural sulfur-containing uridine derivative in tRNA's wobble position, boosts protein translation efficiency/fidelity and enhances RNA stability—key for synthetic RNA therapeutics.

Creative Biolabs offers tailored s²U integration with proprietary, precise modification tech. Backed by insights into s²U's functions, it delivers high-quality RNA tools to accelerate your therapeutic development.

2-Thiouridine Modification Service: Structure, Mechanism, Advantages, Application

Structure and Chemistry

2-Thiouridine (s²U) is a pyrimidine nucleoside where the oxygen atom at the C−2 position of the uridine base is replaced by a sulfur atom. This seemingly minor substitution—the C=O group becoming a C=S group —significantly alters the electronic and steric properties of the nucleoside. The resulting thiocarbonyl group participates in distinct hydrogen-bonding patterns and dramatically influences the ribose sugar conformation.

Mechanism of Action

The key mechanism leveraged by s²U is the conformational locking of the sugar ring. The s²U modification favors the C3'−endo sugar pucker, which is characteristic of the A-form helical structure found in RNA.

• Translational Enhancement: In tRNA, this rigid conformation stabilizes the anticodon stem-loop, improving the fit within the ribosome's A-site and increasing the rate and accuracy of codon-anticodon recognition.
• Structural Stabilization: The sulfur atom enables the formation of an unusual, stabilizing s²U: s²U base pair. This non-canonical pairing significantly contributes to the overall structural robustness of the RNA duplex, providing high thermal stability that resists degradation.

Key Advantages

• Increased Thermal Stability: Prolongs mRNA's functional half-life in manufacturing, storage, and in vivo delivery, ensuring activity when reaching the target site.
• Superior Translation Fidelity: Reduces protein synthesis errors/frame-shifts, guaranteeing full-length functional therapeutic proteins and avoiding non-functional/harmful truncated ones.
• Immune System Modulation: Alters mRNA's molecular shape/recognition profile, reducing innate immune sensor activation to minimize unwanted responses and lower toxicity risks.
• Compatibility and Customization: Works with other modifications (e.g., Ψ) for synergistic effects, enabling customization to specific therapeutic needs and optimizing mRNA performance.

Therapeutic Applications

The Creative Biolabs 2-Thiouridine Modification Service is immediately applicable to next-generation therapeutics:

• mRNA Vaccines: Designing highly stable and efficient vaccine candidates that require lower doses and maintain structural integrity during cold-chain transport.
• Protein Replacement Therapy: Engineering mRNA for systemic administration where sustained, high-level expression of essential proteins (e.g., enzymes) is required.
• Synthetic Biology and RNA Research: Creating chemically defined RNA constructs for in vitro studies, structural analysis (e.g., NMR, crystallography), and fundamental research into RNA replication mechanisms.

Workflow

Required Starting Materials

1. Template Sequence: DNA plasmid or linear PCR template specifying the target gene, UTRs, and Poly(A) tail.
2. Target Cell/Application Data: Info on intended delivery method (e.g., LNP), target cell line, and desired expression kinetics.
3. Desired Purity/Scale: Specific requirements for final RNA quantity (mg to gram scale) and purity level (HPLC required).

Project Initiation & Design

Review client template, define optimal s²U substitution sites and percentage based on therapeutic application (vaccine vs. protein replacement).

Modified Nucleotide Synthesis & QC

Custom synthesizes s²UTP and other non-canonical NTPs, with rigorous QC to ensure >98% purity (critical for high-fidelity IVT).

Optimized In Vitro Transcription (IVT)

Run IVT using customized polymerase mixes and optimized buffers to maximize s²U incorporation efficiency and transcript yield.

Enzymatic Capping & Purification

Apply high-efficiency Cap 1 structure and Poly(A) tailing, followed by automated HPLC purification to remove contaminants and truncated sequences.

Final Quality Control & Release

Conduct comprehensive analysis (capillary electrophoresis for integrity, stability testing for thermal resistance) before client release.

Final Deliverables

1. s²U -Modified mRNA Transcript (mg/g scale): High-purity, capped, tailed final product (lyophilized or in solution).
2. Certificate of Analysis (CoA): Detailed report with mass spectrometry verification of full-length transcript, and qPCR/gel verification of integrity.
3. HPLC Purity Report: Documentation confirming residual DNA template, enzymes, and truncated RNA fragments are below predefined regulatory thresholds.

Estimated Timeframe

6 to 10 Weeks: Duration depends on synthesis scale and complexity of the required modification pattern.

What We Can Offer

The stability and performance of therapeutic mRNA hinge on strategic chemical modification. Creative Biolabs' service is engineered to provide bespoke RNA constructs that deliver superior therapeutic outcomes. We solve key challenges by ensuring your final product features optimal structural integrity and translational efficiency, directly translating to higher protein expression and lower systemic toxicity in vivo.

Scientific Research Testimonials

FAQs

The Creative Biolabs 2-Thiouridine Modification Service represents the pinnacle of precision RNA engineering, providing the enhanced stability and translational functionality required for clinically viable therapeutics. By integrating advanced nucleoside chemistry and rigorous biomanufacturing protocols, we deliver robust, high-purity mRNA that accelerates your program and mitigates critical development risks.

Hot IVT Vectors

Hot IVTScrip™ mRNA Transcript