bioRxiv Science⌕ Search

bioRxiv · 10.1101/2025.11.14.688492

SPrUCE: Utilizing Ultraconserved Elements of DNA for Population-Level Genetic Diversity Estimation

Abstract

Ultraconserved elements (UCEs) provide ideal candidates for targeted sequencing and cost-effective acquisition of genome-wide data. While UCEs have been widely used in phylogenetic studies to recon-struct evolutionary relationships, their use in population-level research has been limited. This limited application stems from uncertainty over whether UCEs can capture the levels of genetic variation needed to answer population genomic questions central to ecology and biodiversity research. The concern is that, by definition, UCEs are highly conserved and may therefore lack sufficient within-species variation. The more variable flanking regions (400-750 bp from the UCE core) contain informative polymorphisms, though diversity decreases near the core. Thus, any naive estimator of genetic diversity that ignores this conservation will have an underestimation bias. In this paper, we introduce SPrUCE: Sigmoid Pi requiring UCEs, a reference-free method that estimates nucleotide diversity{pi} from aligned UCE data. SPrUCE corrects underestimation bias by modeling the change in diversity away from the UCE core using a Gompertz function. The model accounts for the bias introduced by the conserved core and allows for more accurate per-site diversity estimates. We tested SPrUCE on UCE alignments from a range of taxa, including invertebrates and vertebrates (finches, honeybees, sheep, and smelt). SPrUCE produces diversity values consistent with whole-genome derived estimates that require an assembled reference. It is fast, scalable, and effective even with missing data. Its modeling approach enables accurate population-level assessments of genetic diversity, offering a new and reliable option for conservation and population genetics.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Melendez, D., Sapci, A. O. B., Bafna, V., Mirarab, S.. 2025-11-16. SPrUCE: Utilizing Ultraconserved Elements of DNA for Population-Level Genetic Diversity Estimation. https://doi.org/10.1101/2025.11.14.688492

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Integrative Nanopore and Illumina sequencing reveals age-associated tRNA modification and CCA-tail dynamics in yeast

Aging is characterized by a progressive loss of proteostasis. Transfer RNAs (tRNAs) are essential regulators of translation, yet their dynamics during aging remain poorly understood due to challenges in sequencing highly modified RNAs. Here we present a benchmarked Nanopore direct RNA sequencing (RNA004 chemistry) resource that profiles the Saccharomyces cerevisiae tRNAome during replicative aging at single-molecule resolution. Using in vitro transcribed tRNA controls, we establish modification detection thresholds and validate key findings with orthogonal Illumina sequencing. While overall tRNA abundance remains largely stable, our resource reveals age-associated terminal A cleavage at the 3' CCA tail of mature tRNAs, targeted T-loop and anticodon modification changes, and single-molecule evidence of modification co-occurrence. This dataset provides a resource for exploring tRNA regulation, translation fidelity, and longevity.

genomics↗

A hydrogen-producing mitochondrion in an anaerobic eukaryotrophic rhizarian

Diverse eukaryotes thrive under low oxygen conditions, in part through highly modified mitochondrion-related organelles (MROs) that use alternate metabolic pathways to support ATP production and cofactor recycling. Anaerobic lifestyles have evolved repeatedly across the eukaryotic tree of life, each providing an independent opportunity to understand how eukaryotes adapt to life in low oxygen conditions. Here, we use single-cell transcriptomics to reconstruct the MRO metabolism of PCE SSF, a benthic eukaryotrophic flagellate and the first cultivated representative of Novel Clade 12 (NC12; Rhizaria), an independently anaerobic rhizarian lineage. PCE SSF possesses an anaerobic hydrogen-producing mitochondrion capable of hydrogenosome-type substrate-level phosphorylation. It also retains a nearly complete but likely branched tricarboxylic acid pathway that lacks citrate synthase and malate dehydrogenase. The function of citrate synthase may instead be fulfilled by the typically cytosolic ATP citrate lyase, previously reported in this context only in the anaerobic cercozoan, Brevimastigomonas motovehiculus. Unlike B. motovehiculus, however, PCE SSF retains only Complex II and the NuoE/NuoF subunits of the electron transport chain and lacks a mitochondrial genome. Together, these features indicate an atypical and reduced mitochondrial metabolism, highlighting the diversity of evolutionary solutions to anaerobic energy metabolism in eukaryotes.

genomics↗

Targeted CRISPRi screening reveals unexpected resilience across the RNA polymerase III transcriptome

Increased RNA polymerase III (Pol III) activity and tRNA abundance are widely linked to cancer cell growth, yet the functional requirement for individual Pol III genes and core components remains unclear, in part due to the difficulty of achieving gene-specific perturbation of highly conserved loci. Here, we developed an inducible CRISPR interference platform and a custom single-guide RNA (sgRNA) library enabling gene-specific targeting of Pol III-transcribed genes and Pol III machinery. Genome-wide screening identified several Pol III dependencies in diploid fibroblasts and HEK293T cells, including multiple initiator methionine tRNA genes among the strongest fitness dependencies. Unexpectedly, glioblastoma models remained largely insensitive to repression of both individual Pol III genes and core Pol III components, despite efficient target repression. These findings establish a general strategy for gene-specific interrogation of conserved Pol III genes and indicate that glioblastoma models tolerate extensive perturbation of Pol III genes and machinery.

genomics↗