Thermostability: A Growing Need in RNA Biology

For decades, RNA researchers have battled one persistent enemy: ribonucleases (RNases). These ubiquitous enzymes can rapidly degrade RNA, threatening the integrity of downstream assays such as single-cell RNA sequencing (scRNA-seq), reverse transcription quantitative PCR (RT-qPCR), and cDNA synthesis. Traditional protein-based RNase inhibitors have been critical in protecting RNA, but their Achilles’ heel has long been sensitivity to heat and cold-chain reliance.

As molecular workflows become increasingly complex and global supply chains demand more flexibility, the need for thermally robust and glycerol-free RNase inhibitors has grown urgent.

A Recent Breakthrough: SEQURNA in scRNA-Seq

A 2024 study published in Nature Communications introduced SEQURNA, a synthetic thermostable RNase inhibitor tailored for single-cell RNA sequencing workflows (Noble et al., 2024). Researchers demonstrated that SEQURNA:

• Matches or surpasses conventional RNase inhibitors in protecting RNA during scRNA-seq.
• Remains effective through heat-mediated steps, including 72 °C denaturation.
• Reduces cold-chain dependency, enabling simpler logistics and reproducibility.

This marks a pivotal advancement, showing that thermostability in RNase inhibition is no longer just a convenience—it’s becoming a requirement for modern RNA biology.

“The synthetic RNase inhibitor provides additional unique improvements in reproducibility and throughput, enabling new experimental workflows including retained RNase inhibition throughout heat cycles.”
— Noble et al., Nature Communications (2024)

RiboGrip®: Thermostability in Practice

While SEQURNA demonstrates what synthetic RNase inhibitors can achieve, RiboGrip® RNase Inhibitor reflects similar principles already in practice. Designed in silico with a proprietary Stability TAG, RiboGrip® demonstrates remarkable heat resistance:

• Retains full activity after 1 hour at 60 °C.
• Withstands room-temperature storage and shipping for up to one month, alleviating cold-chain constraints.
• Delivered in a glycerol-free, high-concentration (220 U/µl) formulation, making it compatible with lyophilization and air-drying—ideal for regulated environments or field applications.

For experts in molecular biology and diagnostics, this combination of robustness and formulation flexibility is particularly relevant. Glycerol, while useful as a stabilizer, can interfere with sensitive cell-free transcription/translation systems, making a glycerol-free option essential for high-fidelity applications.

Lessons for Expert Users

The convergence of findings from SEQURNA and the demonstrated performance of RiboGrip® points to broader lessons for the field:

1. Thermal Resilience Enables Workflow Expansion
   Heat-stable inhibitors unlock workflows previously limited by enzyme degradation, such as high-temperature denaturation in sequencing protocols.
2. Reducing Cold Chain Dependence Improves Scalability
   Both SEQURNA and RiboGrip® illustrate how reagent stability reduces logistical bottlenecks, especially for global labs or diagnostics firms.
3. Formulation Matters
   Glycerol-free inhibitors not only enable lyophilization but also avoid inhibitory effects in cell-free systems—a growing area of synthetic biology.
4. Regulated Settings Demand Consistency
   IVF centers, molecular diagnostics labs, and QC environments require reagents that can endure varied handling without compromising reproducibility.

The Bigger Picture

The SEQURNA study validates what many in the field have suspected: future RNA workflows will rely on RNase inhibitors engineered for stability, flexibility, and performance under stress. In parallel, tools like RiboGrip® already embody these principles, providing reliable protection for a wide range of applications—from RT-qPCR to RNA-seq to diagnostic assays.

For researchers, lab managers, and procurement specialists, the key takeaway is clear: choosing RNase inhibitors designed with thermal stability in mind isn’t just a convenience—it’s becoming a critical factor in experimental success and logistical efficiency.

Sources

• Noble C.J., et al. Introducing synthetic thermostable RNase inhibitors to single-cell RNA-seq. Nature Communications (2024). Link
• ProLab Supply. RiboGrip® RNase Inhibitor
• Pandi G., et al. Lyophilization of transcription–translation systems enables cell-free synthetic biology. Nature Communications (2022). Link