Reference-free discovery with barcoded single-cell sequencing.

Dehghannasiri, Roozbeh; Kokot, Marek; Starr, Alexander L; Maziarz, Jamie; Gordon, Tal; Tan, Serena Y; Wang, Peter L; Voskoboynik, Ayelet et al. · Nat Biotechnol · 2026

basic_science · Level V

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Abstract

Single-cell RNA sequencing (scRNA-seq) analyses typically focus on alignment and differential gene expression, being blind to other transcriptome variation. Here we present sc-SPLASH for statistics-first, reference-free discovery on barcoded scRNA-seq and spatial transcriptomics; its independent BKC submodule optimizes barcoded data preprocessing and is approximately 50-fold faster than UMI-tools. sc-SPLASH discovers secreted repeat proteins in immune-like cells of sponge (Spongilla; missing from the reference) and tunicate (Ciona).