Biology subjects

Zhao, K.

Publications and source records attributed to Zhao, K..

At least 19 recordsLinked to original sources

bioRxiv2026

LysM: a bacterial self-encoded peptidoglycan hydrolase as an antimicrobial and anti-biofilm agent for Gram-negative nosocomial pathogens

Agents with both anti-biofilm property and broad-spectrum antimicrobial activity are urgently needed, especially for drug-resistant nosocomial pathogens. Here, we report a bacterial genome-encoded glycoside hydrolase, named LysM, with lytic activity against multiple Gram-negative nosocomial pathogens and little toxicity to human cells. 3D structural modelling and site-directed mutagenesis indicates that LysM is a peptidoglycan-specific glycoside hydrolase. In combination with EDTA, exogenous LysM induces explosive bacterial cell lysis, exhibits potent bactericidal activity, and eliminates 24 h-old biofilms formed by Gram-negative ESKAPE group pathogens with the highest activity on three nosocomial pathogens, namely Pseudomonas aeruginosa, Acinetobacter baumannii, and Enterobacter cloacae, all of which contain LysM homologs in their genome. More strikingly, LysM supplied in growth media without EDTA, also inhibits the growth of several nosocomial pathogens. Single-cell tracking analysis reveals that LysM completely prevent biofilm formation of P. aeruginosa by reducing bacterial surface-adherence and extending the doubling time. A LysM-based cocktail consisting of LysM, EDTA, and matrix exopolysaccharide-degrading glycosidase PslG, shows potent anti-biofilm activity against P. aeruginosa, including drug-resistant clinical isolates, both in vitro and in human airway organoids (HAOs) biofilm infection model. In addition, LysM enhances the efficacy of many antibiotics from all major classes, including colistin, tobramycin, ciprofloxacin, and tetracycline. Our findings not only provide a clinical candidate for the treatment of nosocomial infections but also open a new path for antimicrobial discovery and anti-biofilm strategies, as homologues of LysM are present in the genomes of many bacteria and phages.

microbiology↗

bioRxiv2026

Charting sex divergence in the human brain across the lifespan

Biological sex fundamentally shapes human brain organization, but sex-specific normative neuroanatomical trajectories across the lifespan remain largely uncharted. Here, we constructed independent, sex-specific lifespan brain charts using structural neuroimaging data from 59,915 healthy individuals (29,760 males and 30,155 females) ranging in age from 266 postconception days to 100 years. By examining 296 structural phenotypes across global, cortical, and subcortical measures, these models revealed widespread sex differences in maturational timing, with males reaching peak milestones later than females. These trajectories demonstrate that sex differences evolve dynamically, with phenotype-specific windows of emergence and maximal separation. Compared with conventional sex-pooled references, sex-specific models achieved superior predictive accuracy and reduced misestimation of individual deviations in healthy populations. Across five neuropsychiatric conditions, sex-specific models improved the detection of extreme deviations and revealed both shared and sex-dependent patterns of disorder-related neuroanatomical abnormalities. These sex-specific charts establish tailored normative references for assessing brain development, ageing, and disease.

neuroscience↗

bioRxiv2026

Gene functional-state covariance reveals modular structure in the Arabidopsis thaliana dispensable genome

Classical multilocus theory predicts that selection acting on combinations of loci can maintain non-random associations among allelic states, causing selection to act on multilocus genotypes rather than individual alleles. Across eukaryotic species, pangenome studies have documented thousands of genes that vary in functional status between individuals, yet this variation is analyzed as if each gene evolves independently. Here we test for non-random association within the dispensable pangenome across 1,012 natural genotypes of Arabidopsis thaliana to better understand the structure and evolution of eukaryotic pangenomes. Genome-wide pairwise association testing of loss-of-function alleles across genotypes identified 150,033 significantly covarying gene pairs among 5,679 unique genes after controlling for kinship and population structure. Most associations were between genes on different chromosomes, and covarying gene pairs showed greater co-expression and functional similarity than expected by chance. Network analysis revealed a non-random higher-order architecture comprising stable functional communities with distinct biological functions and structured connectivity among communities. The enriched functions and connectivity of communities revealed a tight coupling of modules involving plant immunity, environmental response, and specialized metabolism, highlighting how these systems integrate and evolve in response to dynamic ecological pressures. Taken together, these results show that the dispensable genome has a complex higher-order structure driven by functionally interacting genes, and that variation in gene content provides a population-scale resource for investigating the organization and evolution of biological systems.

genomics↗

bioRxiv2026

trFlow: ultrafast all-atom protein conformational ensemble generation via conditional flow matching

Generating diverse, all-atom protein conformations remains challenging, especially for recovering alternative functional states and modeling conformational ensembles. We present trFlow, an ultrafast approach for generating all-atom conformations using geometry-guided flow matching. trFlow leverages three core innovations: (1) a hybrid single- and multi-step sampling scheme that balances accuracy and speed; (2) a decoupled representation-generation architecture that maximizes computational efficiency; and (3) fine-tuning on NMR ensembles to better capture conformational heterogeneity. Extensive benchmarking across dual-conformation proteins, NMR ensembles, and a CASP16 dynamic target shows that trFlow outperforms competing methods in recovering challenging states and reproducing experimental heterogeneity, while being an order of magnitude faster than existing all-atom approaches. These results establish trFlow as a practical and scalable framework for large-scale generation of all-atom conformational ensembles.

bioinformatics↗

bioRxiv2026

Integrated Transcriptomics and Histological Analysis Reveals Gill Adaptation Mechanisms to Hypoxic Stress in Largemouth Bass (Micropterus salmoides)

Largemouth bass (Micropterus salmoides) is a globally significant freshwater aquaculture species known for its rapid growth, high-quality flesh, and excellent nutritional value. Given the increasing incidence of hypoxia in its aquaculture setting, understanding the molecular mechanisms underlying gill adaptation to hypoxic stress is critical for improving resilience in this species. To achieve this, we employed full-length transcript sequencing using PacBio Iso-Seq and high-throughput Illumina RNA-seq to construct a comprehensive transcriptome profile. Histological analyses revealed apparent structural alterations in gill tissues under hypoxic stress (dissolved oxygen: 2.0 +/- 0.5 mg/L, duration: 96 h), including curved and swollen gill filaments, apoptosis, and epithelial cell vacuolation. Comparative transcriptomic analysis revealed pronounced shifts in the expression of genes involved in energy metabolism, angiogenesis, immune responses, cell proliferation, and apoptosis. Notably, the down-regulation of genes associated with cell cycle and proliferation suggests that M. salmoides may conserve energy by suppressing cell division under hypoxic conditions. In contrast, the increased expression of apoptosis-related genes, including Caspase 7, indicates that apoptosis may play a protective role to maintain tissue homeostasis under hypoxia. Furthermore, the suppression of chemokine-mediated immune pathways implies that hypoxia redirects energy toward essential metabolic processes at the cost of local immune responses. GO and KEGG enrichment analyses identified TNF- and chemokine-mediated signaling pathways as key regulators for tissue homeostasis and adaptive responses to hypoxic stress. These findings enhance our understanding of the mechanisms underlying hypoxia adaptation and provide valuable genetic resources for breeding strategies aimed at enhancing hypoxia tolerance in M. salmoides.

genomics↗

bioRxiv2026

The transcription factor Bhlhe40 promotes inflammatory functions of ILC2s through inducing GM-CSF while inhibiting IL-10 expression

Group 2 innate lymphoid cells (ILC2s) regulate type 2 immune responses partly by recruiting eosinophils, but the mechanisms underlying this process remain incompletely understood. Although both ILC2s and type 2 T helper (Th2) cells are capable of expressing IL-5, a cytokine critical for recruiting eosinophils, ILC2s are more potent than Th2 cells in this process. Here, we show that the transcription factor Bhlhe40 promotes GM-CSF production in ILC2s, and Bhlhe40 expression in ILC2s is required for efficient eosinophil recruitment during both papain- and helminth-induced type 2 immune responses. However, Bhlhe40 deficiency in ILC2s had no effect on the production of classical type 2 cytokines, including IL-4, IL-5 and IL-13, despite Bhlhe40 being required for type 2 cytokine production by Th2 cells. Furthermore, in contrast to ILC2s, Th2 cells produced little GM-CSF and administration of GM-CSF rescued eosinophil recruitment in ILC2-specific Bhlhe40-deficient mice. In addition to promoting GM-CSF expression, Bhlhe40 repressed IL-10 production in ILC2s as it did in Th2 cells, particularly during chronic inflammation. Single-cell transcriptomic analyses further supported this regulatory network, and the ChIP-Seq data revealed direct binding of Bhlhe40 to the enhancer and promoter regions within the Il10 and Csf2 loci. Collectively, our findings show that Bhlhe40 exerts distinct gene regulatory functions in ILC2s and Th2 cells, and Bhlhe40 modulates the inflammatory and anti-inflammatory functions of ILC2s by promoting GM-CSF while suppressing IL-10 production.

immunology↗

bioRxiv2026

Lineage sorting inflates cross-environment functional enrichment

"Everything is everywhere, but the environment selects." What it selects has never been specified: genomes, lineages carrying their inherited gene content, or genes, rewritten inside lineages already in both places. Separating them needs a measurement, not a method: inflation is visible only where uncorrected and corrected estimates sit side by side on the same genes. Comparing only same-genus genomes across six catalogues, the median effect of the 178 orthologues a conventional soil-ocean analysis calls strongly enriched falls from 0.250 to 0.030, 45 reversing sign, DMSP demethylation keeping 2% of its reported effect and high-affinity phosphate transport reversing. On six host-niche contrasts within one Escherichia coli collection the same estimator changes almost nothing, retaining 92 to 100% with no reversals: the collapse belongs to the comparison, not the correction, its size varying over a hundredfold across eight contrasts. Fifty-nine orthologues survive family-wise permutation control, no negative control among them; they concern light, salinity, desiccation and metal efflux, and half are invisible to screening. Re-tested literature claims keep a median 24% of their effect but beat matched random sets 2.7-fold. Reading enrichment as adaptation requires lineage control and a per-contrast bound on what a comparison can manufacture from nothing.

ecology↗

bioRxiv2026

Destabilization of intratumor Tregs by CTLA-4 engagement confers anti-CTLA-4-driven immunotherapy

Immune checkpoint inhibitors (ICI) that are antibodies against CTLA-4 have achieved therapeutic effects on multiple types of cancers 1-4, but the mechanisms underlying the therapy remain incompletely understood. In contrast to the initial theory that the antibody blockades CTLA-4 on T cells is of central importance, it has recently been demonstrated that selective reduction of CD4+Foxp3+ regulatory T cells (Tregs) in the tumor microenvironment by the antibody plays a key role in the anti-tumor effects5-8, although this phenomenon remains under debate in human patients9-12. We show here that anti-CTLA-4 antibody engages CTLA-4 in Tregs specifically in the tumor tissues to reduce their stability and survival and weaken their suppressive function through upregulating TGF-{beta} signaling. This leads to the antibody-mediated - upregulation of CD4+ and CD8+ effector T cell anti-tumor immunity and consequently cancer immunotherapy. Specifically, anti-CTLA-4 antibody directly stimulates CTLA-4 in intratumor Tregs to enhance their TGF-{beta} receptor I (T{beta}RI)-mediated TGF-{beta} signaling13. This results in reduction of IL-2 receptor CD25 expression and downregulation of lactate metabolism in intratumor Tregs to reduce their stability and survival 14-16, and also compromises their suppressive function by inhibiting Foxp3 expression. This activity requires high levels of CTLA-4 expression in the intratumor Tregs and the presence of antibody Fc receptors in the tumor tissues. Strikingly, deletion of T{beta}RI specifically in Tregs completely prevents the reduction of intratumor Tregs and abrogates the anti-CTLA-4-mediated cancer immunotherapy. In contrast to intratumor Tregs, anti-CTLA-4 antibody treatment blocks CTLA-4 in the intratumor CD4+ Foxp3- and CD8+ T effector cells and in the peripheral Tregs to decrease their T{beta}RI expression and increase their expansion due to their relatively low levels of CTLA-4 in the same tumor bearing mice. Significantly, the engagement of CTLA-4 by anti-human CTLA-4 antibody (ipilimumab) also upregulates TGFBR1 but decreases Il2RA expression and lactate metabolism in human Tregs in vitro and in humanized mice in vivo, leading to suppression of tumor progression. We have provided additional mechanism underlying anti-CTLA-4-mediated anti-tumor effects through destabilizing intratumor Tregs by CTLA-4 engagement-mediated TGF-{beta} signaling. This could lay a theoretical foundation for designing optimal immunotherapy based on anti-CTLA-4 antibody in cancer patients.

immunology↗

bioRxiv2026

Hydrogen sulfide dynamically upregulates copper uptake and localization

The reactivity of copper, an essential micronutrient that undergoes facile cycling between Cu1+ and Cu2+ redox states, is carefully controlled within the confines of protein binding sites, and by sequestration in storage vesicles, or harnessed to kill pathogens by active pumping of Cu1+ into phagosomes. We have discovered that hydrogen sulfide, a signaling metabolite generated in copious quantities at the host-microbiome interface, upregulates Cu accumulation in diffusely dispersed puncta across the cell, as visualized by X-ray fluorescence microscopy. The Cu is predominantly in the Cu2+ state with oxygen/nitrogen ligands. Cu import occurs via the non- canonical ZNT1 transporter, while export, following sulfide withdrawal, is ATP7A-dependent. Cu accumulates at the apices of colon crypts in a mouse model of elevated sulfide exposure due to SQOR deficiency in the intestinal epithelium, establishing in vivo relevance. Our study reveals that sulfide is a dynamic regulator of the Cu pool, stimulating Cu2+ influx into highly concentrated puncta.

cell biology↗

bioRxiv2026

Projectomic Organization of the Serotonin System of the Mouse Brain

The serotonin system innervates nearly the entire brain to support diverse functions, and its dysfunction is implicated in multiple psychiatric disorders. Although recent studies suggested that this anatomically diffuse system supports differentiated rather than uniform modulation, its overall organization is unclear. Here, using systematic whole-brain axon tracing and integrated analyses, we show that the serotonin projectome is organized by functional relatedness rather than physical proximity of its targets. Dorsal and median raphe serotonin neurons partition into five projectomic groups, each preferentially innervating the hippocampus, basal ganglia, cortex, medial interbrain, or brainstem/lateral thalamus, with within-group structure ranging from discrete subgroups to continuous variation. Spatial transcriptomic analyses reveal that each projectomic group corresponds to a distinct combination of transcriptomic clusters, with transcriptome and somatic position jointly predicting projectomic identity. Projection-specific serotonin depletion produces dissociable behavioral phenotypes. Together, projectomic identity emerges as a central axis linking axon collateralization, molecular diversity, and behavioral function.

neuroscience↗

bioRxiv2026

Cerebrovascular Single-Nucleus RNA-Seq Reveals Heat Shock Activation and Vascular Remodeling in Alzheimer's Disease and Primary Tauopathies

Cerebrovascular alterations are widely observed in both Alzheimers Disease (AD) and primary tauopathies. Here, we hypothesized that mechanisms of cerebrovascular alterations are shared between AD and primary tauopathies. We performed single-nucleus RNA sequencing of postmortem human inferior temporal gyrus to characterize transcriptomic changes across cerebrovascular cell types in AD and primary tauopathies (Corticobasal Degeneration, Picks disease, and Progressive Supranuclear Palsy). Differential gene expression analyses revealed disease-specific transcriptional programs across vascular cell populations. However, genes involved in the heat-shock response were consistently upregulated across all diseases, suggesting a conserved cerebrovascular stress response during neurodegeneration. We further identified marked cerebrovascular remodeling in AD relative to primary tauopathies, along with dysregulation of genes mapping to AD risk loci in endothelial cells. Transcriptomic findings were validated using tissue clearing, light-sheet microscopy, and immunofluorescence quantification of vascular markers. These results define a conserved vascular stress program alongside AD-specific remodeling, highlighting the vasculature as a therapeutic target in neurodegeneration.

neuroscience↗

bioRxiv2026

Ferrosome Organelles Spatially Insulate a Redox-Active Ferrous Phosphate Biomineral from Cytosolic ROS Chemistry

Ferrosomes are recently discovered lipid-bound bacterial organelles that store iron as iron-phosphate biominerals, yet the chemical nature and physiological consequences of ferrosome-stored iron remain poorly understood. Here, we combined X-ray absorption spectroscopy (XAS), electron microscopy, inductively coupled plasma mass spectrometry (ICP-MS), and physiological analyses to characterize ferrosome iron in Clostridioides difficile. XAS analysis of isolated ferrosomes revealed an amorphous iron-phosphate biomineral containing mixed Fe(II)/Fe(III), consistent with partial oxidation during aerobic isolation. In contrast, whole-cell XAS of intact anaerobically maintained cells demonstrated that ferrosomes predominantly contain a structurally disordered ferrous phosphate biomineral with local Fe-O-P coordination features similar to those of vivianite. Upon air exposure, this ferrous biomineral rapidly oxidized to a ferric phosphate-like state, revealing a highly oxygen-sensitive iron-storage phase. Despite containing abundant redox-active Fe(II), ferrosome-stored iron contributed minimally to the cytosolic labile iron pool. Consistent with this observation, isolated ferrosomes exhibited little ROS-generating activity, and ferrosome-overproducing cells displayed no substantial increase in sensitivity to oxygen, peroxide, or paraquat stress relative to ferrosome-deficient controls. Together, these results establish ferrosomes as iron-storage organelles that sequester redox-active Fe(II) in a mineralized ferrous phosphate phase, limiting its participation in cytosolic ROS chemistry and providing a mechanism for the safe storage of reactive iron. Significance StatementIron is essential for life but can also damage cells because ferrous iron drives oxidative stress. How cells store large amounts of ferrous iron while limiting toxicity therefore remains a fundamental biological question. Ferrosomes are recently discovered bacterial organelles that store iron as iron-phosphate biominerals, but the chemical nature and physiological consequences of ferrosome-associated iron remained unknown. Using Fe K-edge X-ray absorption spectroscopy, we show that ferrosomes in Clostridioides difficile contain a redox-sensitive ferrous phosphate biomineral. Physiological analyses demonstrate that this iron is largely inaccessible to cytosolic reactive oxygen species (ROS) chemistry. These findings reveal that bacteria can combine biomineralization and subcellular compartmentalization to maintain large intracellular iron reservoirs while limiting iron-dependent oxidative damage.

microbiology↗

bioRxiv2026

Ecological determinants of disease and immunity in myelodysplastic syndromes

Myelodysplastic syndromes (MDS) are clonal hematopoietic malignancies characterized by ineffective hematopoiesis, dysplastic morphology, and risk of progression to acute myeloid leukemia. While genomic alterations intrinsic to malignant MDS disease-initiating cells have been well-characterized, molecular assessment of the bone marrow in situ has been limited. Here we present single cell spatial assessment of gene expression, T cell receptors, as well as MDS-defining mutations and RNA isoforms within fixed, decalcified human bone marrow core biopsies (41 MDS, 15 controls) paired with single cell immunogenomic analysis of bone marrow aspirates (35 MDS, 6 controls). Bone marrow spatial analyses of >7.47x106 cells identified hematopoietic and non-hematopoietic populations not readily captured in dissociated tissue. We developed computational methods to compare ecological niche structures, revealing enriched hematopoietic niches and reorganization of T cell immunity in MDS patient bone marrow. In situ genotyping of mutant cells revealed increased TGF{beta} expression in malignant megakaryocytes suppressing local T cell cytotoxicity. By contrast, TGF{beta} signaling was disrupted in mutant cells due to aberrant splicing of multiple TGF{beta} signaling components. This study provides a spatially resolved landscape of human MDS bone marrow and uncovers mechanisms by which malignant cells simultaneously promote intrinsic clonal persistence while rewiring the microenvironment for immune escape.

cancer biology↗

bioRxiv2026

Rhodopsin is a tunable capacitor buffering the toxic, desensitizing retinoids of the vertebrate eye

The eye is a marvel of evolution, but visual sensitivity introduces the risk of blinding photodamage. Here we reveal that the locus of sensitivity--the visual pigment rhodopsin --moonlights as a tunable mechanism of retinal photoprotection. Independent of signaling, light activated rhodopsin (R*) serves as an overflow capacitor buffering all-trans retinal (atRAL), a toxic and desensitizing retinoid agonist that accumulates as lipofuscin--a clinical marker of macular degeneration. Across mammals, R* stability reflects binding affinity and sequestration of atRAL (capacitance), thereby mitigating phototoxicity to a degree proportional to species photodamage risk, with human R* uniquely non-protective. A mouse model of defective atRAL clearance treated with a synthetic R* of unnaturally high atRAL capacitance preserved retinal function following light damage. This gene therapy also provided supra-physiological scotopic sensitivity despite being a signal-silent receptor, shielding neighboring dark-state receptors from agonist interference during regeneration, while also enhancing the signaling of endogenous R*. While the consensus human rhodopsin is not photoprotective, during recent evolution, mutations that protect against light damage emerged in high irradiance human environments and are now significantly associated with a 38% reduced risk of blindness and low vision. Together, our findings redefine rhodopsin as a tunable light buffer that can be leveraged to enhance photoreceptor function beyond natural limits.

biochemistry↗

bioRxiv2026

Zero-shot design of a de novo metalloenzyme

The de novo design of enzymes remains a central challenge, requiring consideration of catalytic mechanism and optimization across biochemical and biophysical criteria. Here we present dEVA (design by EVolutionary Algorithm), a multi-objective protein design framework built on principles drawn from evolutionary biology. We apply dEVA to the zero-shot, de novo design of metalloenzymes by optimizing the coordination sphere of catalytic metals. We characterize a bizinc metalloenzyme that exhibits promiscuous hydrolytic activity towards both phosphomonoesters and phosphodiesters. This design achieves a rate enhancement ((kcat/KM)/kw) up to 3 x 1013, comparable to characterized natural phosphatases. dEVA offers a general and modular strategy for the programmable design of protein function without dependence on natural templates or evolutionary information.

biochemistry↗

bioRxiv2026

TriGraphQA: a triple graph learning framework for model quality assessment of protein complexes

Accurate quality assessment of predicted protein-protein complex structures remains a major challenge. Existing graph-based quality assessment methods often treat the entire complex as a homogeneous graph, which obscures the physical distinction between intra-chain folding stability and inter-chain binding specificity. In this study, we introduce TriGraphQA, a novel triple graph learning framework designed for model quality assessment of protein complexes. TriGraphQA explicitly decouples monomeric and interfacial representations by constructing three geometric views: two residue-node graphs capturing the local folding environments of individual chains, and a dedicated contact-node graph representing the binding interface. Crucially, we propose an interface context aggregation module to project context-rich embeddings from the monomers onto the interface, effectively fusing multi-scale structural features. We conducted comprehensive tests on several challenging benchmark datasets, including Dimer50, DBM55-AF2, and HAF2. The results show that TriGraphQA significantly outperforms state-of-the-art single-model methods. TriGraphQA consistently achieves the highest global scoring correlations and lower top-ranking losses. Consequently, TriGraphQA provides a powerful evaluation tool for protein-protein docking, facilitating the reliable identification of near-native assemblies in large-scale structural modeling and molecular recognition studies.

bioinformatics↗

bioRxiv2026

Structural basis of receptor retro-translocation in peroxisomal protein import

Peroxisomes import all matrix proteins post-translationally from the cytosol, a process that requires recycling of cargo receptors across the peroxisomal membrane. The membrane-embedded ubiquitin ligase, composed of Pex2, Pex10, and Pex12, is central to this process, but its mechanism remains unclear. Here we determined cryo-electron microscopy structures of the Saccharomyces cerevisiae Pex2-10-12 complex in closed and open states bound to Pex8, an essential factor of previously undefined function. The structures reveal how Pex2-10-12 gates its retro-translocation pore to control receptor entry and how the closed-to-open transition repositions the Pex10 RING domain to enable receptor mono-ubiquitination. Pex8 docks onto Pex2-10-12 from the matrix and guides receptors into the pore. Functional analyses show that the receptors N-terminal segment downstream of its mono-ubiquitination site initiates a loop insertion into the pore. These findings establish how Pex2-10-12 coordinates receptor recognition, retro-translocation, and ubiquitination, providing the molecular basis for receptor recycling in peroxisomal protein import.

biochemistry↗

bioRxiv2026

Ryder: Epigenome normalization using a two-tier model and internal reference regions

Motivation: Sequencing-based epigenomic profiling methods are powerful but suffer from technical variability that complicates cross-sample comparisons and can obscure true biological signals. While existing normalization methods using spike-in controls or computational approaches have been proposed, they often rely on assumptions that may not hold across diverse experimental conditions or require additional data types. Results: We present Ryder, a flexible and robust Python package for the normalization of epigenomic signal tracks. Ryder leverages stable internal reference regions, such as invariant CTCF binding sites, to correct for technical artifacts genome-wide. Our results show that it effectively adjusts both background noise and signal intensity, ensuring accurate signal alignment across samples while preserving genuine biological differences. We demonstrate that Ryder performs robustly across diverse assays including DNase-seq, CUT&RUN, ATAC-seq, MNase-seq, and ChIP-seq, with or without spike-in controls. By reducing technical noise, Ryder improves the detection of genuine biological changes, such as quantitative reduction of chromatin accessibility at key enhancer elements by depletion of BRG1, a key subunit of the chromatin remodeling BAF complexes. Availability and Implementation: The Ryder source code, documentation and test data are freely available at: https://github.com/YaqiangCao/ryder . The software version used in this study is archived at Zenodo: https://zenodo.org/records/21267457 .

bioinformatics↗