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Biology subjects

Jiang, W.

Publications and source records attributed to Jiang, W..

At least 37 records · Page 2Linked to original sources

Explainable Physicochemical Determinants of Protein Ligand Binding via Non-Covalent Interactions

Protein-ligand binding governs enzymatic catalysis, metabolic homeostasis, and therapeutic modulation. Thus, the accurate prediction of these interactions underpins modern rational drug discovery. However, existing deep-learning frameworks largely operate as black-box predictors that fail to resolve the individual residues mediating binding or decode the fundamental non-covalent forces that drive molecular recognition. To address these limitations, we present ExplainBind, an interaction-aware framework that predicts binding likelihood, localizes specific binding residues at single-amino-acid resolution rather than coarse pocket-level regions, and decodes the underlying non-covalent interaction patterns, all zero-shot, without requiring prior three-dimensional structural inputs. To support residue- and interaction-level training and evaluation, we construct InteractBind, a protein-ligand benchmark with residue-atom interaction maps. Simultaneously, benchmarking experiments demonstrate that ExplainBind consistently outperforms state-of-the-art baselines across diverse protein and ligand spaces, maintaining high precision when generalized to entirely novel sequences and chemical scaffolds. When applied to two unseen therapeutic targets, ExplainBind successfully ranks potent angiotensin-converting enzyme (ACE) inhibitors and clarifies differences in their potency via affinity-stratified interaction landscapes. Furthermore, we demonstrate the prospective utility of ExplainBind by discovering novel inhibitors and activators of L-2-hydroxyglutarate dehydrogenase (L2HGDH) through wet-lab validation, with mechanistically distinct interaction profiles providing a clear molecular rationale for their divergent functional outcomes. Collectively, these results establish ExplainBind as a powerful, generalizable tool for mechanistically informed, interpretable drug discovery.

molecular biology↗

rRNA Expansion Segments Mediate Ribosome Dimerization as a Conserved Stress Response

Inhibition of mRNA translation is a common feature in proteostatic stress cellular responses. Puromycin, a widely used compound for studying translation, disrupts protein synthesis by mimicking the 3 end of aminoacyl-tRNAs. Despite its extensive use as a research tool to probe the connection between translation activity and various physiological and pathological states, the cellular response associated with puromycin-induced translation stress remains incompletely understood. Utilizing in situ cellular electron tomography and topology analysis, we visualized the translation machinery at high resolution. Our analysis revealed that puromycin-treated neuronal cells exhibit an accumulation of "idle ribosomes" characterized by the binding of eIF5A, indicating a close association of this factor with translationally inactive ribosome states under stress. Additionally, the idle ribosomes formed dimeric complexes mediated by ribosomal RNA expansion segments, suggesting an evolved mechanism involving these regions in translation hibernating and protecting idle ribosomes. We show that the hibernating disome formation is not unique to puromycin administration but represents a conserved mechanism as a response to different cellular stressors including those associated with endoplasmic reticulum (ER) stress and amino acid depletion. Altogether, our findings shed light on previously unexplored aspects regarding unique states of mammalian ribosome hibernation, and collectively offers new avenues for understanding the correlation of cellular stress response and the regulation of protein synthesis.

molecular biology↗

AbNovoBench: a resource and benchmarking platform for monoclonal antibody de novo sequencing

Monoclonal antibodies (mAbs) are critical in disease diagnostics and therapeutics, yet the performance of mass spectrometry (MS)-based de novo sequencing remains incompletely characterized due to limited antibody-specific datasets and the absence of a standardized benchmark framework. Here we present AbNovoBench, a comprehensive framework for evaluating data analysis strategies for mAb de novo sequencing. It features the largest high-quality dataset to date, generated in-house, comprising 1,638,248 peptide-spectrum matches from 131 mAbs across six species and 11 proteases, supplemented by eight mAbs with known full-length sequence for end-to-end reconstruction assessment. Employing a unified training dataset, we systematically benchmarked 13 deep learning-based de novo peptide sequencing algorithms and three assembly strategies across peptide sequencing metrics (accuracy, robustness, efficiency, error types) and assembly metrics (coverage depth, assembly score). AbNovoBench (https://abnovobench.com) provides an online platform enriched with curated antibody MS resources and pre-trained models, enabling customizable antibody sequencing workflows, accelerating antibody-specific algorithms development, and improving reproducibility in proteomics.

systems biology↗

UBA1 Mitigates Myocardial Ischemia/Reperfusion Injury by Attenuating Endoplasmic Reticulum-Mitochondria Contacts via Pdzd8 ubiquitination

BACKGROUNDMyocardial ischemia/reperfusion injury (I/RI) represents a serious clinical complication in patients after acute myocardial infarction. Ubiquitin-activating enzyme 1 (UBA1) catalyzes the initial step of ubiquitination and plays a fundamental role in regulating protein homeostasis and related diseases. This study aims to elucidate the functional contribution of UBA1 to the pathogenesis of myocardial I/RI and to uncover its underlying mechanisms. METHODSSingle-cell RNA sequencing was employed to characterize UBA1 expression in human ischemic heart tissues. Myocardial I/R injury was examined in myocardial-specific UBA1 knockout (UBA1cko) mice, UBA1-overexpressing mice (rAAV9-UBA1), and corresponding controls. Neonatal rat cardiomyocytes underwent hypoxia/reoxygenation in vitro. Cardiac function and infarction were evaluated by echocardiography and pathological staining. Protein-protein interactions were analyzed via immunoprecipitation combined with mass spectrometry. The endoplasmic reticulum-mitochondrial contact sites (ERMCSs) and mitochondrial ultrastructure were evaluated through transmission electron microscopy and confocal imaging. RESULTSUBA1 expression was significantly downregulated in human and murine ischemic myocardium, especially in cardiomyocytes. UBA1cko mice exhibited aggravated I/RI with greater infarct size, impaired function, apoptosis, elevated intracellular Ca2+ levels, mitochondrial dysfunction, and ER stress, whereas UBA1 overexpression conferred cardioprotective effects. Mechanistically, UBA1 directly bound to and ubiquitinated Pdzd8, a key ERMCS-tethering protein, thereby promoting its degradation, which inhibited ERMCS formation and improved mitochondrial dysfunction and ER stress. Moreover, knockdown of Pdzd8 via rAAV9-siRNA effectively mitigated UBA1 knockout-induced myocardial damage. Additionally, administration of auranofin (AF), a U.S. Food and Drug Administration-approved drug for treating rheumatoid arthritis, markedly alleviated myocardial I/RI via activating UBA1 in vivo and in vitro. CONCLUSIONSUBA1 confers protection against myocardial I/RI by limiting ERMCS formation through Pdzd8 ubiquitination. Activating UBA1 or targeting Pdzd8 as a potential therapeutic strategy for the treatment of ischemic heart disease. GRAPHIC ABSTRACTA graphic abstract is available for this article. Clinical PerspectiveO_ST_ABSWhat Is New?C_ST_ABSO_LIUBA1 expression is downregulated in human and murine ischemic myocardium, especially in cardiomyocytes. C_LIO_LICardiac deletion of UBA1 significantly exacerbates myocardial ischemia/reperfusion injury (I/RI), whereas cardiac UBA1 overexpression confers a marked protective effect. C_LIO_LIUBA1 interacts with Pdzd8 (PDZ domain containing 8) and facilitates its ubiquitination and subsequent degradation, which then reduces endoplasmic reticulum-mitochondria contact sites (ERMCSs) and ameliorates mitochondrial dysfunction and ER stress, protecting myocardial I/RI. C_LIO_LIPharmacological activation of UBA1 with the FDA-approved drug auranofin attenuates myocardial I/R injury and improves heart dysfunction. C_LI What Are the Clinical Implications?O_LIUBA1 represents a new therapeutic target for myocardial I/RI. C_LIO_LIActivating UBA1 or targeting Pdzd8 may offer a promising therapeutic strategy for mitigating myocardial I/RI and heart failure, underscoring its potential for clinical translation. C_LI

pathology↗

Disrupted TRiC allostery and conformational equilibrium by LCA-linked mutations: structural and functional insights

The eukaryotic chaperonin TRiC/CCT is essential for proteostatis, yet the molecular basis of its subunit-specific pathologies remains poorly understood. Here, we elucidate the molecular mechanism of Leber Congenital Amaurosis (LCA), a severe hereditary retinal dystrophy arised from mutations in the CCT2 subunit of TRiC. By integrating cryo-electron microscopy, biochemistry, and proteomics, we demonstrate that LCA-associated mutations (T400P and R516H in CCT2) disrupt TRiCs critical intra-molecular and intra-ring allosteric network and impair its functional cycle, drastically reducing the population of the folding-active closed state. Unexpectedly, we captured a fully folded endogenous -tubulin within the mutant TRiC chamber, revealing a unique CCT8 C-terminal tail involved folding pathway, distinct from {beta}-tubulin. Furthermore, cellular proteomics revealed that TRiC dysfunction causes a specific downregulation of essential SLC membrane transporters. We propose that the loss of these transporters is likely catastrophic for metabolically demanding tissues like the retina and developing embryo. Our work provides a direct mechanistic link between TRiC structural defects and LCA pathology, offering a new framework for understanding etiology of chaperonopathies.

biochemistry↗

Efflux Pump Activation Confers Mupirocin Resistance and Enhances Rhizosphere Fitness in Pseudomonas

Mupirocin (Mup) is a polyketide antibiotic for clinical MRSA treatment and agricultural biocontrol, acting by binding to IleRS to inhibit protein synthesis. Here, we demonstrate that Pseudomonas viciae 11K1 acquires resistance to mupirocin not through canonical mutations in ileS, the gene encoding the drug target, but via single nucleotide polymorphisms (SNPs) of EmhR, a transcriptional repressor. These SNPs attenuate EmhRs DNA-binding ability, resulting in derepression of the resistance-nodulation-division (RND) efflux pump EmhABC. This leads to a 7-fold (to 800 g/mL) increase in the mupirocin minimum inhibitory concentration (MIC), and confers cross-resistance to multiple other antibiotics. Critically, 11K1 strains harboring EmhR mutations exhibit enhanced competitive fitness in colonizing wheat rhizospheres against the mupirocin-producing Pseudomonas bijieensis 2P24. The conservation of this regulatory mechanism in Pseudomonas aeruginosa, mediated by the EmhR ortholog NalD, underscores its broader biological significance. Our findings establish a direct link between efflux pump regulation and ecological adaptation, highlighting a key mechanism contributing to environmental antimicrobial resistance with important implications for the clinical and agricultural use of mupirocin. IMPORTANCEMupirocin and its producing Pseudomonas strains are widely used in both clinical and agricultural contexts, making mupirocin resistance a significant concern for public health and food security. While canonical mupirocin resistance is primarily attributed to mutations in ileS, the gene encoding the drug target isoleucyl-tRNA synthetase (IleRS), our study identifies a novel resistance mechanism in Pseudomonas viciae strain 11K1 mediated by SNPs in the transcriptional repressor EmhR. These SNPs derepress the RND efflux system EmhABC, and reduce intracellular mupirocin levels. This mechanism not only enhances tolerance to mupirocin but also confers cross-resistance to multiple antimicrobial agents, raising the risk of multidrug-resistant strain spread. EmhR-mutant 11K1 strains exhibit enhanced rhizosphere competitiveness in the presence of mupirocin-producing Pseudomonas bijieensis strain 2P24, indicating an ecological fitness advantage that could promote resistant population expansion in agriculture environments. The same regulatory pathway is conserved in human pathogen Pseudomonas aeruginosa PAO1 via the functional ortholog NalD, suggesting that this mechanism may be widespread in Pseudomonas. These findings fill a critical gap in understanding non-target-based mupirocin resistance, clarify the ecological drivers of antimicrobial resistance (AMR), and offer practical insight for improving mupirocin application to limit the emergence and spread of resistant pathogens.

microbiology↗

Two phytohormones synergistically induce parasitic weeds seed germination via KAI2d receptors

Root parasitic plants can severely decrease global crop production. The germination of their seeds is induced by host root-derived strigolactones (SLs). Considering this unique germination system, a "suicidal germination" method has been proposed to control root parasitic plants. However, this method requires the cost-effective production of germination inducers. In this study, we determined that jasmonates and SLs can synergistically induce the seed germination of root parasitic plants (Orobanche minor and Striga hermonthica). Biochemical analyses indicated that jasmonates and SLs cooperatively activate multiple divergent KARRIKIN INSENSITIVE 2 receptors. Our findings have elucidated the host recognition systems associated with these highly duplicated promiscuous receptors, with potential implications for developing a new strategy to protect crops against root parasitic plants.

plant biology↗

Computer-assisted dissection of the DNA mismatch recognition mechanism of the MutS protein

By sensing local conformational strains in DNA, TaqMutS utilizes a push-pull mechanism via the synergy of residues S57 and D472 to modulate {pi}-stacking interactions between the key recognition residue F39 and DNA, thereby achieving specific mismatch-site recognition. Meanwhile, residues K61 and R473 fine-tune non-specific binding of TaqMutS through electrostatic synergy.

synthetic biology↗

Female-predominant anti-CD4 IgG autoantibody production and its correlations with plasma levels of progesterone, microbial translocation, and blunted immune reconstitution in HIV-infected patients on suppressive ART

Autoimmunity contributes to HIV immunopathogenesis even in the absence of overt autoimmune disease. We previously showed that anti-CD4 autoantibodies from people with HIV (PWH) on suppressive antiretroviral therapy (ART) can mediate cytotoxicity against CD4+ T cells, implicating a role in impaired immune reconstitution. Despite viral suppression, many PWH with poor CD4 recovery exhibit chronic immune activation, microbial translocation, and dysregulated humoral immunity. Here, we identify a female-predominant elevation of plasma anti-CD4 IgG autoantibodies in aviremic PWH receiving ART. Across two independent cohorts, HIV-positive females, but not males, displayed significantly higher anti-CD4 IgG, predominantly IgG1, compared with HIV-negative controls, without parallel increases in anti-CD4 IgA or IgM. This sex-specific pattern was unique to anti-CD4 IgG and was not observed for anti-CD8 IgG, anti-double-stranded DNA IgG, or anti-nuclear antigen IgG; these control autoantibodies correlated with one another but not with anti-CD4 IgG. Elevated anti-CD4 IgG levels were associated with lower plasma progesterone levels and reduced absolute CD4+ T-cell counts. Markers of microbial translocation, soluble CD14 (sCD14), lipopolysaccharide-binding protein (LBP), and lipopolysaccharide (LPS), were also selectively increased in HIV-positive females, with sCD14 and LBP showing significant or borderline associations with anti-CD4 IgG. Together, these findings identify anti-CD4 IgG as a sex-dimorphic autoimmune signature in treated HIV infection, linked to progesterone levels, persistent microbial translocation, and incomplete immune recovery. This work highlights an under-recognized intersection of sex, mucosal barrier dysfunction, and autoimmunity in HIV pathogenesis and suggests potential therapeutic targets to improve immune reconstitution in women.

pathology↗

Brainstem GLP-1 neurons modulate physiological satiation and drive sustained weight loss in obese mice

Glucagon-like peptide-1 receptor (GLP-1R) activation in the brain strongly reduces appetite, but most brain GLP-1Rs are not accessible for systemically administered GLP-1R agonists. Acute activation of nucleus tractus solitarius (NTS) GLP-1 neurons, targeting brain GLP-1Rs, strongly suppresses food intake separate from GLP-1R agonists. However, it is unknown if their chronic stimulation is a viable strategy for appetite suppression, or if obesity disrupts their function. Here we demonstrate that GLP-1 neurons, distributed through NTS and IRT, determine meal size and their number is inversely correlated with bodyweight gain. GLP-1 neurons in IRT and NTS differ in their inputs but generate outputs in largely overlapping areas. Higher body weight and fat mass predicts a higher percentage of active GLP-1 neurons. Their chemogenetic activation is an efficient means of reducing food intake, and in obese mice chronic activation elicits sustained weight loss. In conclusion, GLP-1 neurons are a feasible target for obesity treatment.

neuroscience↗

Cannabis-enriched oral Actinomyces induces anxiety-like behavior via impairing mitochondria and GABA signaling

The human oral microbiome is increasingly recognized as a contributor to brain health, yet its mechanisms remain unclear. Our previous work revealed that oral Actinomyces species was enriched in chronic cannabis smokers. Here, we show oral inoculation of cannabis use-associated Actinomyces species, especially A. meyeri, to wild-type C57BL/6 mice leads to anxiety-like behaviors, non-region-specific microglia activation, mitochondrial dysfunction, and reduced GABAergic neurotransmission, without evidence of bacterial translocation to the brain, neuroinflammation, and memory decline. Notably, Actinomyces species-producing metabolites, i.e., arginine and argininosuccinate, were increased in both oral swabs and brain following inoculation in vivo. These Actinomyces species-producing metabolites induced mitochondrial dysfunction and oxidative stress in neurons in vitro, indicating a neuropathogenic role and aligning with reduced GABAergic neurotransmission in vivo. Together, these results suggest that oral cannabis-associated dysbiosis impacts behavior through mitochondrial stress and impaired inhibitory signaling, indicating the oral-brain metabolic axis is potentially consequential in neuropsychiatric disorders. TeaserChronic heavy cannabis use-enriched oral bacteria can drive anxiety and neuropathogenesis in mice. Highlights{whitebullet} Cannabis-associated oral Actinomyces enrichment induces anxiety-like behavior in mice {whitebullet}Microglial activation occurs without neuroinflammation (IL-1{beta}, TNF-, and IL-6) {whitebullet}Mitochondrial hyperactivation and reduced inhibitory GABAergic signaling Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=160 SRC="FIGDIR/small/689724v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@120b0b1org.highwire.dtl.DTLVardef@1304782org.highwire.dtl.DTLVardef@a6aa77org.highwire.dtl.DTLVardef@17d6d_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

ZONAB regulates DNA methylation and mitochondrial function to control endothelial cell senescence

Regulation of the endothelial stress response is important for blood vessel homeostasis and angiogenesis, processes disrupted in common vascular diseases and ageing. Here, we discovered that the Y-box factor ZONAB (ZO-1-associated nucleic acid binding protein; YBX3), a gene associated with risk loci for severe vascular disorders, regulates endothelial homeostasis and angiogenesis. By combining cell-based assays with primary endothelial cells and genome-wide expression and methylation measurements, we found that ZONAB depletion results in mitochondrial deregulation, increased reactive oxygen species and a defective oxidative stress response, as well as increased promoter methylation of cell cycle genes. Consequently, ZONAB depletion triggered cellular senescence via a phosphatidylinositol 3-kinase (PI3K)/Akt-dependent pathway, which was attenuated by an antioxidant or by drugs targeting mitochondrial function or fragmentation. Thus, our results reveal how ZONAB controls changes in gene expression and DNA methylation to regulate endothelial proliferation, inflammation, and angiogenesis, indicating a central role of ZONAB in vascular health.

cell biology↗

Flexible parental care in a songbird correlates with sex-specific responses to seasonal phenology, mating opportunity and reproductive success

O_LISpecies with diverse parental care patterns often exhibit flexibility in response to environmental changes. Such changes can influence parental care decisions by altering the trade-offs between current and future reproduction. While previous studies have revealed correlations between specific social and ecological factors and parental care patterns, the joint effects of multiple factors could be rather complex and may result in different outcomes within and across populations, and between sexes. C_LIO_LIIn this study, we revealed parental care systems and their seasonal variations across three geographic populations in a polygamous species, the Chinese penduline tits (Remiz consobrinus), by monitoring breeding events over years. To investigate how local conditions (including social and ecological conditions) correlate with sex-specific parental care patterns, we compared breeding phenology and local conditions (including season length, mating opportunity and reproductive success) across the populations. C_LIO_LIStriking population differences in parental care strategies are found in this species: biparental care predominated in one population, whereas female-only care was more common in the other two. Our analyses reveal that, despite a large difference in season length, the time of egg-laying peaked similarly across the three populations. Female penduline tits determined the breeding phenology, which varies male mating opportunities dramatically over the season. For all populations, male Chinese penduline tits were more likely to desert the clutch when male mating opportunities were high. In contrast, females consistently provided care, regardless of variation in female mating opportunities. C_LIO_LIWe also found that fitness costs on the current brood resulting from offspring desertion differed across populations. This desertion cost on brood fitness matters more than achieving a high reproductive reward in driving the emergence of uniparental care, contrary to the theory that uniparental care only occurs when single parents can efficiently raise nestlings alone. C_LIO_LIOur study demonstrates that similar parental care patterns across populations could result from different underlying factor interactions and highlights the consistent sex-specific responses to local conditions in parental care. C_LI

ecology↗

Characterize Oral-to-Blood Microbial DNA Translocation in Individuals with Cocaine Use Disorder

BackgroundCocaine disrupts gut barriers in animal models, potentially enabling microbial translocation and inflammation in the periphery and central nervous system (CNS), but its direct role in inducing inflammation remains controversial. This study aimed to determine if the oral cavity is a source of circulating microbial DNA translocation in individuals with current cocaine use disorder (CUD). ResultsA cross-sectional case-control study was conducted, comparing CUD and demographically matched non-drug controls. Ten CUD (via smoking or vaping) and 24 controls provided paired saliva and blood samples. Microbial 16S rRNA V4 region was sequenced in isolated microbial DNA from saliva and plasma. Single-cell RNA sequencing (scRNAseq) was analyzed in human peripheral blood mononuclear cells. Saliva from CUD, but not plasma, exhibited reduced alpha diversity and altered beta diversity, characterized by enriched Streptococcus and depleted Fusobacterium, Neisseria, and other taxa relative to controls. Controls exhibited low to undetectable microbial translocation in plasma. By contrast, plasma displayed CUD-specific oral enrichment of several Streptococcal species and evidence of translocation into the bloodstream. S. parasanguinis, but not cocaine alone, induced IL-1{beta} and TNF- production in human primary monocytes in vitro. scRNAseq further revealed innate immune activation, impaired T cell function, and heightened susceptibility to infection in CUD. ConclusionsThis pilot study demonstrating that CUD via smoking or snorting exhibited oral microbial dysbiosis and selective oral-to-blood microbial translocation in vivo. These findings suggest that a compromised oral-to-blood barrier, rather than cocaine itself, promotes immune perturbations in CUD.

microbiology↗

A spatially-resolved human brain proteome atlas for understanding function and disease

While the brain performs specialized functions across distinct regions, the spatial organization of the human brain proteome remains largely uncharted. Here we present a comprehensive spatially-resolved proteome atlas of the human brain, analyzing over two thousand MRI- guided locations across four individuals. Proteome analysis integrated with transcriptomics reveals extensive post-transcriptional regulation, with cortical regions showing markedly higher protein diversity than transcript. Unsupervised molecular clustering defines distinct brain territories that transcend anatomical boundaries, instead reflecting metabolic demands and functional specialization patterns. Application to epilepsy brain tissue uncovered disrupted astrocyte metabolism, protein homeostasis and therapeutic targets including the seizure-associated purinergic receptor P2RX7. This resource bridges molecular and systems neuroscience to accelerate neurological drug discovery.

neuroscience↗

The Implications of Alternative Splicing Regulation for Maximum Lifespan

Mammalian maximum lifespan (MLS) varies over a hundred-fold, yet the molecular mechanisms underlying this diversity remain unclear. We present a cross-species analysis of alternative splicing (AS) across six tissues in 26 mammals, identifying hundreds of conserved AS events significantly associated with MLS, with the brain containing twice as many tissue-specific events as peripheral tissues. MLS-AS events are enriched in pathways related to mRNA processing, stress response, neuronal functions, and epigenetic regulation, and are largely distinct from genes whose expression correlates with MLS, indicating that AS captures unique lifespan-related signals. The brain exhibits certain associations divergent from peripheral tissues and reduced overlap with body mass (BM)-associated splicing; neither is observed at the gene expression level. While MLS- and age-associated AS events show limited overlap, the shared events are enriched in intrinsically disordered protein regions, suggesting a role in protein flexibility and stress adaptability. Furthermore, MLS-associated AS events display stronger RNA-binding protein (RBP) motif coordination than age-associated ones, highlighting a more genetically programmed adaptation for lifespan determination, in contrast to the more variable splicing changes seen with chronological aging. These findings suggest alternative splicing as a distinct, transcription-independent axis of lifespan regulation, offering new insights into the molecular basis of longevity.

bioinformatics↗

Composing trajectories for rapid inference of navigational goals

Animals efficiently learn to navigate their environment. In the laboratory, naive mice explore their environment via highly structured trajectories and can learn to localize new spatial targets in as few as a handful of trials. It is unclear how such efficient learning is possible, since existing computational models of spatial navigation require far more experience to achieve comparable performance and do not attempt to explain the evolving structure of animal behavior during learning. To inform a new algorithm for rapid learning of navigational goals, we took inspiration from the reliable structure of behavior as mice learned to intercept hidden spatial targets. We designed agents that generate behavioral trajectories by controlling the speed and angular velocity of smooth path segments between anchor points. To rapidly learn good anchors, we use Bayesian inference on the history of rewarded and unrewarded trajectories to infer the probability that an anchor will be successful, and active sampling to trim hypothesized anchors. Agents learn within tens of trials to generate compact trajectories that intercept a target, capturing the evolution of behavioral structure and matching the upper limits of learning efficiency observed in mice. We further show that this algorithm can explain how mice avoid obstacles and rapidly adapt to target switches. Finally, we show that this framework naturally encompasses both egocentric and allocentric strategies for navigation.

animal behavior and cognition↗

Characterization and engineering of highly efficient Cas12j genome editors

The large size of CRISPR-Cas enzymes limits their delivery for therapeutic applications. Cas12j nucleases offers hypercompact alternative but show moderate editing efficiency. To overcome this limitation, we identified eight novel Cas12j orthologues (Cas12j-11 to Cas12j-18) from viral metagenomes. All showed low editing activity in mammalian cells. We engineered T5 exonuclease-Cas12j fusions (T5Exo-Cas12j), two of which, T5Exo-Cas12j-12, and -18 exhibited up to 42% editing in HEK293T and 9% in K-562 cells, outperforming wild-type Cas12j counterparts and comparable to LbCas12a. Intriguingly, robust in cellula editing in both HEK293T and K-562 cells was strictly dependent on the presence of 5'-TAC trinucleotides within the target DNA sequence. Furthermore, we fused the Cas12j orthologues with the TadA8e deaminase and developed base editors, termed Be-(d)Cas12j. Among these, Be-(d)Cas12j-13 demonstrated efficient A-to-G base conversion in mammalian cells. This study expands the CRISPR toolbox by characterizing and engineering novel Cas12j orthologues into compact, high-efficiency genome editors.

bioengineering↗