SEQURNA enhances FLASH-seq gene detection while eliminating DTT dependence

This dataset contains single-cell RNA sequencing data from a systematic benchmarking study of SEQURNA, a synthetic thermostable RNase inhibitor, against commercial recombinant RNase inhibitors (RRIs) using the FLASH-seq protocol. The data are provided as an integrated AnnData object (.h5ad) containing gene expression counts, cell-level quality metrics, and annotations for all cells passing quality control.

We hypothesized that SEQURNA could replace conventional protein-based RNase inhibitors in full-length single-cell RNA sequencing while eliminating the requirement for dithiothreitol (DTT), a reducing agent routinely added to stabilize commercial RRIs. To test this, we processed human retinal organoid cells and peripheral blood mononuclear cells (PBMCs) using FLASH-seq across multiple 384-well plates, varying SEQURNA concentration (0.2, 0.5, 0.75, 1, 1.25, and 1.5 U/µl) and DTT supplementation (0, 5, and 10 mM). Control conditions included Takara RRI, Watchmaker Genomics RRI, DIANA RRI, and no-inhibitor controls. Retinal organoids represent a moderate-to-high RNA content cell type, while PBMCs represent a challenging low-RNA model. A subset of retinal organoid samples was stored at −80°C for one month prior to processing to assess storage stability. Data were generated on two sequencing platforms (Illumina NovaSeq and Element AVITI) to assess cross-platform reproducibility.