Introducing synthetic thermostable RNase inhibitors to single-cell RNA-seq

Joyce Carol Noble, Antonio Lentini, Michael Hagemann-Jensen, Rickard Sandberg and Björn Reinius ()
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Joyce Carol Noble: Karolinska Institutet
Antonio Lentini: Karolinska Institutet
Michael Hagemann-Jensen: Karolinska Institutet
Rickard Sandberg: Karolinska Institutet
Björn Reinius: Karolinska Institutet

Nature Communications, 2024, vol. 15, issue 1, 1-9

Abstract: Abstract Single-cell RNA-sequencing (scRNAseq) is revolutionizing biomedicine, propelled by advances in methodology, ease of use, and cost reduction of library preparation. Over the past decade, there have been remarkable technical improvements in most aspects of single-cell transcriptomics. Yet, little to no progress has been made in advancing RNase inhibition despite maintained RNA integrity being critical during cell collection, storage, and cDNA library generation. Here, we demonstrate that a synthetic thermostable RNase inhibitor (SEQURNA) yields single-cell libraries of equal or superior quality compared to ubiquitously used protein-based recombinant RNase inhibitors (RRIs). Importantly, the synthetic RNase inhibitor provides additional unique improvements in reproducibility and throughput, enables new experimental workflows including retained RNase inhibition throughout heat cycles, and can reduce the need for dry-ice transports. In summary, replacing RRIs represents a substantial advancement in the field of single-cell transcriptomics.

Date: 2024
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DOI: 10.1038/s41467-024-52717-4

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