ALT Optical pooled screens connect genetic perturbations to changes in cell morphology and behavior, but turning the microscopy data into interpretable results takes complex, multi-day analyses. Established pipelines process this data, yet researchers still select parameters, inspect outputs, and resolve failures for each experiment. Here, we present Brieflow-auto, a pipeline-specific harness implemented as a Claude Code plugin that lets a coding large language model (LLM) operate Brieflow, an open-source end-to-end pipeline for optical pooled screening data. A researcher supplies experimental facts and approves the launch. An agent tunes parameters in an interactive notebook, and the harness validates the configuration, monitors execution, recovers from common failures, and checks output quality. For pipeline defects, the agent opens upstream issues proposing backward-compatible fixes for the broader community. In five previously analyzed screens, harness runs yielded more cells with a co

ALT Comprehensively characterizing cellular diversity is critical for understanding tissue development, identifying disease-associated populations and investigating gene programmes linked to specialized functions. Advances in single-cell profiling reveal additional cell-specific transcripts and molecular associations, yet many foundation models represent only a restricted portion of this increasingly rich molecular information. Here we present CellNet, which enables panoramic modelling of single-cell transcriptomes by integrating DNA-derived gene identities and measured RNA expression in a shared image representation. Pretrained on 231 million cell profiles spanning 40 species, CellNet improves rare-population and condition-associated state recognition in evaluated cohorts while retaining competitive general cell representations. It predicts withheld coding-gene expression and selected non-coding targets in total-RNA data, and recovers lineage-specific expression patterns in spatial measur

ALT Predicting single-cell perturbation responses requires mapping between two unpaired populations of cells measured before and after the perturbation. Optimal transport (OT) provides a principled framework for constructing such mappings by minimizing the transport cost, and neural OT methods (e.g., CellOT) have been employed for this prediction task. However, these methods rely on a fixed transport cost, typically the squared Euclidean distance. This cost may connect cells with the wrong responses. Lineage tracing provides the information needed to learn the cost, as heritable barcodes link the cells of the same clone before and after the perturbation. Here, we propose CloneOT, a neural inverse OT method that learns both the transport cost and the transport map from partially lineage-traced single-cell datasets. CloneOT{ rep}resents the cost as a neural network. It learns the cost by minimizing the extra cost that the lineage adds to the optimal transport, so that the cheapest transport

ALT Is a chromosome a miniature of its genome, with the same mix of functional elements as the whole? We ask this of eight annotated genomes: human, mouse, zebrafish, Caenorhabditis elegans, Drosophila melanogaster, Saccharomyces cerevisiae, Arabidopsis thaliana and maize. Elements are classified in a four-level hierarchy, from five major classes (protein-coding genes, ncRNA genes, transposable elements (TEs), pseudogenes, IG/TR gene segments) down to finer classes. Units are chromosomes or arms, cytogenetic bands, equal cuts of each chromosome, and random segments of matching size. We compare each unit with the genome and with other units, call compositions homogeneous when they differ by less than 5%, and score each against a random sample of the same size. The first finding is a surprising homogeneity. The level-1 compositions of the major chromosomes match each other and the genome in maize, zebrafish, yeast and C. elegans, and in fly down to the major arms. Finer classes can be homoge

ALT Over 90% of disease-associated variants from genome-wide association studies fall in noncoding regulatory regions, yet their functional interpretation remains a central open problem in genomic medicine. Large language models prompted to interpret such variants routinely hallucinate transcription factor (TF) binding changes, fabricate experimental support, and assign biological significance to statistically negligible signals. We present ARGUS (Agentic Regulatory Genomics for an Uncertainty-aware Scientist), which strictly separates deterministic biological computation from LLM-mediated reasoning. ARGUS wraps 458 DNABERT-based TF binding models in a hypothesis-directed investigation loop where a planner selects evidence sources based on current uncertainty, a verifier deterministically interprets each observation, and intermediate results change the investigation path. On variant rs6983267 at the 8q24 cancer risk locus, the same planner produces four divergent trajectories for four TFs.

ALT Single-cell RNA-sequencing analyses predominantly use either Seurat (R) or Scanpy (Python), with the choice often driven by programming language preference. However, by default, these tools produce divergent variable features, neighbour graphs, clusters, and marker genes. Laboratories requiring Python, which supports most deep-learning, foundation-model, and agent tools, must re-implement Seurat analyses in a framework that does not replicate Seurat's results. Truecell was developed as a Python implementation of the Seurat interface, preserving function names, arguments, default settings, and the object model to facilitate seamless transfer of analyses. In eighteen paired end-to-end evaluations against R Seurat, deterministic outputs matched to floating-point precision, and fold-change order was identical across all nine differential expression tests. In a three-arm benchmark on three datasets, with fixed user parameters and 20 seeds per tool, Truecell more closely reproduced Seurat's

ALT Predicting transcriptional responses to genetic perturbations is central to understanding gene function. Existing predictors primarily rely on transcriptomic measurements, although chromatin accessibility provides complementary information about the cellular context in which perturbations act. Using this information requires linking chromatin context to specific perturbations and accounting for baseline differences between independently sampled control and perturbed populations. We propose ChromaPert, a chromatin-informed framework for predicting state-dependent perturbation responses. ChromaPert combines molecular and DNA-informed locus priors with paired control RNA–ATAC features to jointly represent target identity and measured cellular context. Its Chromatin-Guided Response Router (CGRR) builds a source bank of control-derived expression offsets and retrieves relevant sources using perturbation/locus or control-ATAC similarity. These offsets account for baseline differences, while

ALT Motivation: Plant disease resistance genes, particularly those encoding NB-LRR proteins, are important targets for crop improvement. Proteome-based domain or motif searches can only identify NB-LRRs among existing gene models, meaning they cannot recover loci that have been missed or incorrectly predicted by the reference annotation. The full-length, homology-based R-gene prediction (HRP) method circumvents this issue by reconstructing gene models directly on the genome. However, the original implementation of this method requires several separate phases of classification, comparison and filtering operations. Results: HRPv2 is an automated, enhanced version of the original strategy. The labour-intensive, step-by-step curation process used in HRP has been replaced by a reproducible filtering framework. Enhancing two steps of the homology search process has enabled HRPv2 to better account for the specific NB-LRR variability of the genome. Performance validation confirmed that the number

ALT Genomic language models can write synthetic DNA that carries no record of its origin. A generation-time watermark could provide a provenance signal, if a verifier can detect it using only the DNA sequence, a key, and published detector settings, without knowing where the generated region starts, which strand it lies on, or how it was divided into six-base tokens. We embed the SynthID tournament watermark in two genomic language models, Carbon and GENERator-v2. The verifier searches both strands, every start position, and four window lengths, and corrects its decision for the whole search. In a detection cohort of 1,544 held-out prompts, it found all 3,088 marked sequences per model, unedited and after one substituted, inserted, or deleted base. For ordinary sequences, the one-sided 95% upper confidence bound on the false-positive rate was at most 0.850%; one bound for the wrong-key control reached 1.015% (Carbon, after a deletion). In a development cohort, neither model showed a detect

ALT Genome-wide association studies identify genomic variants associated with traits. Mixed linear model association (MLMA) methods using a whole-genome relationship matrix, such as those implemented in GCTA, are powerful but computationally expensive. Here, we remove key memory and CPU bottlenecks in GCTA, reducing REML memory usage by nearly 75% and substantially accelerating MLMA by orders of magnitude in biobank-scale cohorts while preserving exactness. We further exploit relatedness in the mapping cohort through a reduced-rank Woodbury matrix approach, delivering further orders of magnitude performance gains with controlled genomic inflation. Native on-the-fly dominance recoding also eliminates slow I/O-operations on intermediate files, enabling efficient additive and dominance MLMA analyses in large agricultural cohorts.

ALT Reference genome assemblies are foundational resources across the biological sciences as they begin to expose the fundamental building blocks of each species and the molecular toolkits vital for adaptation and survival. High-quality genomes enable insights into evolution, facilitate ecosystem monitoring and protection and conservation of species, access to new biomaterials and biomedicine, advances in agri- and aquaculture, and support planetary health. The amount of information gathered from a species' genome assembly is directly dependent on its quality. However, different sequencing technologies and software solutions generate a wide range of quality outcomes. Here, the Earth BioGenome Project's Sequencing and Assembly committee, together with a large community of researchers performing eukaryote sequencing and assembly, outlines quality standards for a "reference" assembly and formulates recommendations to ensure these standards are met.

ALT Predicting phenotypic and transcriptional responses to perturbations at single-cell resolution provides a powerful tool for probing biological systems. However, existing methods typically rely on fixed mappings learned during training, making it challenging to calibrate distribution shifts or adapt to novel perturbation conditions during inference. Here, we present scEGFlow, an energy-guided flow matching framework that dynamically bridges control and perturbed cellular states. scEGFlow models continuous transitions from control cell populations to perturbed states using conditional flow matching. It then applies condition-specific energy gradients to correct and steer these predictions, enabling flexible adjustments without retraining the flow model. Evaluations across benchmarks spanning imaging phenotypes and transcriptomic profiles show that scEGFlow outperforms existing methods in reconstructing response distributions under both seen and unseen perturbation conditions, faithfully

ALT The extensive diversity of viral genomes challenges alignment-free representations to capture both nucleotide composition and the higher-order organization of recurring sequence patterns. Conventional k-mer representations quantify pattern abundance but do not explicitly encode how these patterns are organized across scales. Here, we introduce CANN, an alignment-free framework that combines persistent commutative-algebraic descriptors with neural representation learning for viral family classification. CANN uses persistent facet descriptors to encode the multiscale organization of k-mer patterns and complementary k-mer counts to capture sequence composition. Convolutional and Transformer encoders independently learn latent representations of these features, and cosine distances within their respective embedding spaces are averaged before k-nearest-neighbor classification. Across multiple NCBI viral genome collections, CANN achieves robust family-level performance, with the fusion of en

ALT K-mers, short subsequences of length k, play an important role in computational analyses using biological sequence data. A critical downstream processing step involves getting back to the source sequences of selected k-mers for further analysis or validation. Here, we present MerKurio, a high-performance command line tool written in Rust with two main functionalities: 1) extracting sequence records from FASTA/FASTQ files using k-mers and 2) annotating or filtering aligned sequences in SAM/BAM files with k-mer tags. The tool automatically selects the pattern matching algorithm via empirically derived rules based on query characteristics and supports paired-end reads, compressed input files, and reverse complement searching. Benchmark comparisons demonstrate that MerKurio outperforms existing tools in terms of speed while providing additional utility including detailed matching statistics and comprehensive file format support. MerKurio is a fast and user-friendly tool for sequence record

ALT Single-cell transcriptomics enables profiling of cellular states at unprecedented resolution, but its high dimensionality, sparsity, and technical batch effects pose significant challenges for representation learning. Existing single-cell foundation models typically encode each cell independently or only model cells from the same batch for denoising, thereby underutilizing the rich relational information across batches and cell types to model gene expression patterns. We argue that single-cell models can benefit from more informative cell-context modeling. By comparing consistency and variation across cells, models can capture fine-grained gene-gene dependencies associated with cell states, which are essential for learning high-quality representations. Inspired by the use of multiple sequence alignment (MSA) context in protein modeling, we propose CellMSA, a single-cell representation learning framework that introduces an MSA-inspired inductive bias into transcriptomic modeling. For ea

ALT Accurate classification of viral families is essential to understanding genomic diversity, but rapid evolution and heterogeneous genome structure complicate reliable assignment. Newly discovered viruses, particularly those from underrepresented ecosystems, expand the known diversity of viral genomes. This growing diversity poses challenges for classification methods that rely on similarity to existing references. We present Fragment Path Complex Networks (FPCN), a framework that represents viral genomes as sequences of ordered fragments at various scales. FPCN captures contextual sequence information within each fragment and uses path complexes to model complex relationships between neighboring fragments. We evaluated FPCN on well-established NCBI benchmarks, and FPCN achieved higher classification accuracy than competing methods. Further evaluations on newly released datasets revealed FPCN's ability to generalize to genomes that were not presented in the training. Together, these resu

ALT Single-cell cancer datasets are predominantly cross-sectional and rarely provide paired or longitudinal observations linking individual healthy-like cells to tumor-associated states. We introduce CancerZigZag, a seed-initialized diffusion-based framework for exploratory generation of tumor-associated single-cell candidate clouds from unpaired epithelial cell populations. For each cancer context, a diffusion model is trained exclusively on tumor-derived epithelial cells and applied through repeated partial latent-space perturbation and reverse diffusion to held-out healthy-like seeds, generating stochastic candidate clouds without paired measurements or classifier guidance. We applied CancerZigZag to colorectal, breast, lung, and renal cell carcinoma contexts and explored parameter landscapes defined by perturbation depth and the number of ZigZag cycles. Across the reported operating configurations, candidate clouds contained outputs classified toward held-out tumor-derived reference po

ALT Many DNA foundation models are pretrained by masking parts of a sequence and asking the model to reconstruct them. Standard masked pretraining exposes the encoder to special [MASK] tokens that are absent at inference, creating a mismatch between training and downstream use. The role of an explicit global sequence representation such as a [CLS] token and how it should be trained also remain poorly understood for DNA barcodes. We introduce BarcodeMAE+ and study model architecture, global [CLS] representation, and auxiliary pretraining objectives across arthropod COI (BIOSCAN-5M) and fungal ITS (UNITE+INSD) barcodes. Across both barcode regions, the encoder-decoder MAE-LM architecture outperforms its matched encoder-only counterpart in nearly all evaluated configurations, supporting MAE-LM as an effective architectural design for DNA barcode foundation models. A trained global [CLS] representation provides substantial additional gains: on BIOSCAN-5M, [CLS] accuracy increases from 47.53% w

ALT Summary: Functional enrichment analysis (FEA) is a widely used approach for interpreting high-throughput omics data. However, essential methodological details, such as software versions, analysis parameters, and annotation database releases among others, are often incompletely reported, limiting the reproducibility and transparency of enrichment analyses and complicating the assessment of potentially problematic methodological choices. Here we present EMMA, an R/Bioconductor package that integrates with existing FEA tools and automatically captures provenance metadata, such as annotation metadata, software version, and parameters, during the analysis runtime. Our package provides utilities for accessing and exporting the recorded metadata to facilitate transparent reporting and preserve provenance required for reproducible enrichment analyses. This also enables auditing of the results while remaining compatible with existing Bioconductor workflows. Availability and implementation: EMMA

ALT Tokenization is a central design choice in genomic language models, yet most deoxyribonucleic acid (DNA) tokenizers use characters, fixed-length k-mers, or frequency-derived subwords without explicitly using prior information about the specificity of DNA-binding regulatory factors. We introduce Motif-Vocab, a biologically informed tokenizer that scans both DNA strands for statistically calibrated motif matches, emits transcription-factor (TF) identity tokens, and applies nucleotide, k-mer, or byte-pair encoding (BPE) to unmatched sequence. Motif-specific null distributions put position-weight matrices (PWMs) of different lengths and degeneracy on a common significance scale; deterministic overlap rules make the representation reproducible. In controlled Bidirectional Encoder Representations from Transformers (BERT) pretraining on two billion base pairs, real motif libraries outperform randomized-motif controls on 54 of 55 in-scope downstream tasks. On a motif-disjoint recognition task