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bioRxiv · 10.64898/2025.12.12.693841

SEQURNA enhances FLASH-seq gene detection while eliminating DTT dependence

Khven, I.·Ribeiro, M. M.·Crausaz, S.·Picelli, S.
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Abstract

Effective RNase inhibition is critical for single-cell RNA-sequencing, yet commercial recombinant RNase inhibitors (RRIs) require reducing agents for stability and impose substantial costs. Here, we systematically benchmark SEQURNA, a synthetic thermostable RNase inhibitor, against commercial alternatives using FLASH-seq in human retinal organoids and peripheral blood mononuclear cells (PBMCs). SEQURNA at 0.5-1 U/l achieved 14-50% higher gene detection than Takara RRI, with the greatest improvement in low-RNA PBMCs. Surprisingly, DTT supplementation at standard concentrations (5-10 mM) significantly impaired gene detection across all SEQURNA concentrations without improving RNA quality metrics, challenging established reverse transcription protocols. SEQURNA preserved biological heterogeneity, maintained sample stability during one-month storage at -80{degrees}C, and reduced reagent costs by 70%. We recommend SEQURNA at 1 U/l without DTT as an optimized formulation that simultaneously enhances data quality and cost-effectiveness for full-length single-cell RNA sequencing.

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Khven, I., Ribeiro, M. M., Crausaz, S., Picelli, S.. 2025-12-15. SEQURNA enhances FLASH-seq gene detection while eliminating DTT dependence. https://doi.org/10.64898/2025.12.12.693841

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