Biology subjects

Tian, H.

Publications and source records attributed to Tian, H..

At least 19 recordsLinked to original sources

bioRxiv2026

Using a lanthanide tagged nanobody to visualise Glial fibrillary acidic protein by secondary ion mass spectrometry

Lanthanide-labelled antibodies have been widely used for multiplex imaging of proteins directly from tissue using two approaches, imaging mass cytometry (IMC) and variants of secondary ion mass spectrometry (SIMS). Here we show the same approach can be taken using SIMS, but utilising lanthanide-labelled nanobodies instead. Nanobodies offer several advantages over conventional antibodies for antigen tagging, owing to their small size (12-15 kDa), high stability, and solubility. Their reduced molecular size enables greater diffusion through tissue, facilitating access to target sites and allowing them to reach regions that may be less accessible to larger antibodies. This enhanced accessibility can promote more effective localisation and increase binding to target antigens. In the work presented here, we produced a lanthanide-tagged nanobody and assessed its efficacy and compatibility with mass spectrometry imaging (MSI). We showed successful detection of Glial fibrillary acidic protein (GFAP) in tissue by correlating MSI of a lanthanide-tagged anti-GFAP nanobody with immunofluorescence microscopy imaging (IF). The use of nanobodies is an excellent alternative to current antibody tagging methods for SIMS and IMC.

molecular biology↗

bioRxiv2026

Super-Resolution Visual ProteomEx for Hard Tissues and Clinical Samples

Correlating nanoscale tissue architecture with unbiased molecular composition within the same specimen remains a fundamental challenge in spatial biology. Current spatial proteomics methods either lack the imaging resolution required to resolve nanoscale tissue architecture or achieve only limited proteome coverage, and untargeted approaches capable of deep protein identification have yet to be integrated with super-resolution imaging within a single workflow. Here, we present microProteomEx, an integrated platform that combines hydrogel-assisted tissue expansion with mass spectrometry-compatible fluorescence imaging, laser capture microdissection, and bottom-up proteomics to achieve simultaneous three-dimensional super-resolution imaging (effective lateral resolution down to [~]47 nm on conventional diffraction-limited microscopes) and spatially resolved proteomics at a scalable lateral resolution of 29-100 {micro}m (0.02-0.28 nL volumetric resolution). By optimizing fixation, protein anchoring, and a secondary re-embedding strategy, we extend the method to mechanically resilient tissues, including mouse kidney and heart, as well as to formalin-fixed, paraffin-embedded clinical specimens. We demonstrate single-glomerulus and single-plaque proteomics (450-1,000 proteins per structure), resolve proteomic differences between malignant melanoma and giant congenital melanocytic nevus in a rare pediatric case, and characterize the morphology-resolved molecular heterogeneity of cored versus diffuse amyloid-beta plaques in a mouse model of Alzheimers disease across two disease stages. microProteomEx establishes a broadly accessible framework for correlating tissue ultrastructure with deep spatial proteomics, with direct implications for disease biology, biomarker discovery, and precision medicine.

biochemistry↗

bioRxiv2026

STN-DBS frequency-tuned beta subband dynamics are associated with speech tempo in Parkinson's disease

In Parkinsons disease, subthalamic nucleus deep brain stimulation (STN-DBS) frequency modulates narrow beta subbands, with low-frequency stimulation (LFS) selectively attenuating 15-23 Hz power. The negative association between this subband and speech tempo, together with the LFS-related improvement in articulatory rate, suggests that 15-23 Hz subband dynamics may serve as a candidate neural marker associated with speech tempo modulation.

neuroscience↗

bioRxiv2026

LipoTag: A minimal motif for live and functional imaging of plant cell membranes.

The plant plasma membrane is a highly dynamic structure that is crucial for cell compartmentalization, the maintenance of (bio)chemical gradients, signaling and cell growth and responses to stress. In plants, plasma membranes are tightly connected to the cell walls that encase them. These cell walls can act as diffusion barriers and prevent the use of a wide range of synthetic fluorescent probes that have been developed to study animal cell membranes, which lack a cell wall, with live functional imaging. Here, we introduce LipoTag, a minimal chemical motif that, upon chemical conjugation, transforms hydrophobic fluorophores into water-soluble, membrane-targeted probes that can permeate plant cell walls to reach their intended location. LipoTag uses a localized positive charge in combination with a short aliphatic spacer to direct cargo to the plasma membrane. We used LipoTag to develop a suite of membrane-specific fluorescent probes that work in walled organisms beyond the plant kingdom. In addition, we used LipoTag to develop functional reporters for the quantitative imaging of membrane density, lipid order and membrane oxidation in living plant tissues. LipoTag forms a modular platform for exploring the plant plasma membrane with a suite of contemporary imaging modalities.

plant biology↗

bioRxiv2026

Coronavirus envelope protein drives iron sensing disorder by hijacking the TAp73-FDXR axis

Iron overload is increasingly recognized as a critical contributor to coronavirus pathogenesis1, yet the underlying induction mechanisms remain unclear. Here, we uncover a fundamental pathway by which coronavirus drives IRP1 RNA-binding activity to induce iron accumulation2 via targeting the TAp73-FDXR axis. Specifically, coronavirus infection represses transcription of FDXR (encoding the key rate-limiting enzyme in host iron-sulfur cluster synthesis3), thereby impairing host iron-sulfur cluster generation to trigger the functional conversion of the cytosolic aconitase 1 (ACO1) into iron-regulatory protein 1 (IRP1)4, ultimately leading to the hosts persistently false perception of iron deficiency. We identify TAp73 as the primary transcription factor governing FDXR expression, and demonstrate that the coronavirus envelope protein (CoV-E) orchestrates TAp73 nuclear export. Subsequently, CoV-E binds TAp73 through a critical valine residue within its C-terminal PBM domain, inducing the K48-linked ubiquitination and proteasomal degradation of TAp73. Furthermore, we developed a CoV-E-targeting molecule, DPTP-FC, which blocks CoV-E-TAp73 interaction via forming steric hindrance and effectively alleviates iron accumulation and tissue damage caused by PEDV, PDCoV, and SARS-CoV-2 infection. Our study reveals the central role of the TAp73-FDXR axis in CoV-induced iron accumulation, highlighting CoV-E as an attractive antiviral target and DPTP-FC as a promising therapeutic candidate.

microbiology↗

bioRxiv2026

Gut Microbiota Dysbiosis-Mediated Gut NLRP3 Inflammasome Activation Exacerbates Corticospinal Tract Injury After Intracerebral Hemorrhage

Intracerebral hemorrhage (ICH) causes severe neurological deficits, largely attributable to corticospinal tract (CST) injury. However, the underlying mechanism remains unclear, which hinders the development of effective treatment methods. Here, we found that CST injury is not only associated with the activation of NLRP3 inflammasome in the surrounding area of the hematoma as expected, and dysfunction of the blood-brain barrier, but also related to the severe imbalance of the gut microbiota, the activation of NLRP3 inflammasome in the gut and the impairment of gut barrier function after ICH. We therefore systematically investigated how the gut NLRP3 inflammasome and gut dysbiosis exacerbate CST injury after ICH. Knockdown of colon NLRP3 significantly attenuated CST injury and downregulated inflammasome signaling in both the peripheral circulation and the peri-hematomal brain. Consistently, antibiotic-mediated gut microbiota depletion suppressed NLRP3 activation in the gut and brain, improved neurological function, and reduced CST damage. Crucially, fecal microbiota transplantation (FMT) from ICH donors established that the exacerbation of CST injury is dependent on gut dysbiosis. While FMT induced severe pathology in control mice, this effect was improved in gut NLRP3 knockdown recipients, demonstrating that gut NLRP3 is essential for mediating the harmful effects of microbiota dysbiosis. Our findings described a causal gut-brain axis in ICH, wherein microbiota dysbiosis activates the gut NLRP3 inflammasome to exacerbate CST injury, thereby identifying the gut NLRP3 inflammasome as a promising therapeutic target.

neuroscience↗

bioRxiv2026

A structurally unique effector shared between Verticillium dahliae and Fusarium oxysporum is involved in cotton defoliation and virulence on other hosts

Defoliating (D) strains of the vascular wilt fungus Verticillium dahliae cause severe yield losses in cotton and olive worldwide, yet the genetic basis underlying this pathotype has remained unknown. Here, we combined comparative genomics, functional genetics, structural analysis, and phylogenomics to uncover the molecular determinant of defoliation. We identified a small, D pathotype-specific genomic region encoding two duplicated secreted effector genes. Simultaneous deletion of both gene copies abolished pathogenicity and defoliation in cotton, olive, Nicotiana benthamiana, and Arabidopsis thaliana, whereas single deletions reduced virulence and genetic complementation restored disease symptoms. Conversely, expression of the D effector in non-defoliating strains was sufficient to induce defoliation. Moreover, or exogenous application of the purified protein, induced wilting and defoliation as well. Structural analyses revealed that D homologs share a conserved but previously uncharacterized protein fold and are distributed across Verticillium and Fusarium species, exhibiting functional diversification and host-specific activity. Phylogenomic and genomic context analyses indicate repeated horizontal transfer events mediated by giant transposable elements known as Starships. Together, our findings identify the D effector as a central pathogenicity factor that drives defoliation and virulence, and demonstrate how Starship-mediated horizontal gene transfer shapes the emergence and dissemination of an agriculturally devastating fungal trait.

plant biology↗

bioRxiv2026

SLECA: a single-cell atlas of systemic lupus erythematosus enabling rare cell discovery using graph transformer

Systemic lupus erythematosus (SLE) is a highly heterogeneous autoimmune disease with complex immune and molecular dysregulation. While rare immune cell populations are increasingly recognized as critical drivers of disease pathogenesis and progression, the lack of sufficiently powered, comprehensive single-cell atlases has limited their systematic identification and characterization. To address this gap, we present SLECA, the first large-scale single-cell atlas of SLE, enabled by a novel graph-transformer framework for the interpretable discovery and analysis of disease-relevant rare cell populations. SLECA integrates 366 single-cell samples with standardized clinical and biological metadata, providing a comprehensive and analytically unified atlas of systemic lupus erythematosus. By enabling scalable integration and interpretable analysis, SLECA resolves 54 distinct cell types, including rare populations with critical disease relevance. Notably, we identify double-negative T cells (DNTs) as a disease-expanded population whose abundance correlates with clinical severity. Through in silico perturbation, we demonstrate that key transcription factors, specifically JUN and EGR1, can reprogram DNT cells toward conventional T-cell phenotypes, highlighting actionable regulatory vulnerabilities in SLE.

bioinformatics↗

bioRxiv2026

Protein engineering of a genetically encoded biosensor for wastewater detection of profen NSAIDs

Non-steroidal anti-inflammatory drugs (NSAIDs) are pervasive environmental contaminants due to their frequent and widespread use, multiple paths of release into surface and ground water supply, diversity of the chemical class, and toxicity to aquatic and other non-target species. In particular, the 2-arylpropionic acid ("profen") class of NSAIDs poses significant risks to aquatic ecosystems due to incomplete removal during wastewater treatment. Current monitoring precludes high frequency testing at point sources. Here we present the engineering and application of a genetically encodable, protein-based biosensor for the detection of the NSAIDs ketoprofen and pranoprofen in wastewater effluent. We repurposed the plant hormone receptor PYR1 to bind selectively to profens using computational protein design, deep mutational scanning, and yeast 2 hybrid and yeast surface display screening. The resulting sensor, PYRNSAID, has a nanomolar limit of detection for ketoprofen and panoprofen, and {micro}M sensitivity to the NSAIDs ibuprofen, fenoprofen, tolmetin and diclofenac. We also demonstrated dose responsive-activity of our sensor in simulated wastewater matrices containing the common wastewater contaminants sulfamethoxazole, caffeine, acetaminophen, and 2,4, dichlorophenol using a split Nanoluc luminescence assay. PYRNSAID is the first step towards a scalable, cost-effective alternative for real-time monitoring of pharmaceutical pollution.

bioengineering↗

bioRxiv2026

Ac-FAPI-04 Promotes Anti-tumor Immunity via IL-27 Activation and Tnfaip3 Inhibition

BackgroundAccumulating evidence substantiates the capacity of targeted radionuclide therapy (TRT) to potentiate tumor immunotherapy responses. However, combining TRT and immunotherapies exhibits substantial varying efficacy, underscoring the importance of investigating mechanisms through which TRT remodels the tumor microenvironment (TME). Despite recent advancements, the specific signaling molecules mediated by TRT that regulate the functions of cytotoxic T lymphocytes (CTLs) remain unclear. MethodsGiven the demonstrated potential of -radionuclides targeting the fibroblast activation protein (FAP) in TRT, this study employed [225Ac]Ac-FAPI-04 to investigate the mechanisms of TRT-induced TME remodeling. The cytotoxic effects were first evaluated in the B16F10 cell line, followed by an assessment of its in vivo therapeutic efficacy in tumor-bearing mice. RNA-sequencing analysis was utilized to elucidate underlying signaling pathways involving immune responses. Results[225Ac]Ac-FAPI-04 significantly induced DNA double-strand breaks, further elevated ROS levels, and apoptosis in tumor cells. Transcriptomic profiling revealed that [225Ac]Ac-FAPI-04 activates the IL-27/JAK-STAT pathway to amplify interferon-mediated responses, indicating IL-27 serves as a critical regulatory signal for CTLs during TRT. A concomitant observation was the downregulation of Tnfaip3 following [225Ac]Ac-FAPI-04 treatment. Tnfaip3 is implicated in modulating CTLs behavior and intratumoral infiltration, while its downregulation was associated with the upregulation of cytotoxic effector molecules, as confirmed in this study. ConclusionThis study identifies a TRT-induced pattern of synergistic signaling mediated by IL-27 and Tnfaip3, which collectively enhances CTL recruitment and potentiates their cytotoxic function. These findings provide a detailed mechanistic rationale for strategies aimed at augmenting the therapeutic efficacy of TRT and immunotherapy combinations.

molecular biology↗

bioRxiv2025

An inherent T cell-activating mRNA delivery platform for in vivo CAR T generation

The clinical success of CAR T cell therapy has underscored the need for scalable, non-invasive strategies for in vivo T cell engineering. Although mRNA delivery offers a promising alternative, current lipid-based vectors lack intrinsic T cell tropism and often rely on antibody conjugation, complicating manufacturing and raising safety concerns. Here we present an inherent T cell-activating polymer-lipid nanoparticle (ERTLNPs) that enable ligand-free, efficient mRNA transfection and activation of T cells in vivo. ERTLNPs, composed of p-toluenesulfonyl arginine (RT)-functionalized polyethylenimine (denoted as PEI-RT) and helper lipids, preferentially accumulate in the spleen following systemic administration. Without exogenous stimulation, ERTLNPs intrinsically activate T cells, triggering robust mRNA expression and proliferation. Mechanistically, ERTLNPs engage the PI3K/AKT/mTOR signaling axis to reprogram T cell metabolism, promoting expansion while restraining exhaustion. Systemic delivery of mRNA encoding fibroblast activation protein chimeric antigen receptor (mFAP CAR) via ERTLNPs leads to in situ generation of functional CAR T cells, which efficiently eliminate pathological fibroblasts in models of cancer and fibrosis, with minimal off-target effects. This ligand-free, metabolically reprogramming mRNA delivery platform provides a clinically translatable approach for in vivo CAR T cell generation and broadens the landscape of non-viral immunotherapeutic engineering.

bioengineering↗

bioRxiv2025

Gut metabolite IPA alleviates white matter injury post-ICH by enhancing myelin debris phagocytosis via Stap1 inhibition

BackgroundIntracerebral hemorrhage (ICH) causes neurological dysfunction and white matter injury (WMI) characterized by myelin loss, axonal injury and myelin debris accumulation. Microglia-mediated debris clearance is critical for WMI repair. The microbiota-gut-brain axis plays an essential role in the central nervous system diseases, one of the ways in which gut microbiota affects brain is via producing metabolites. Indole-3-proprionic acid (IPA), a tryptophan-derived metabolite that mainly produced by Clostridium sporogenes, exhibits anti-inflammatory and neuroprotective properties. However, its effect on ICH remains unclear. This study aims to investigate the IPA level after ICH and the therapeutic effects of IPA on neurological deficits and WMI, as well as the potential mechanisms underlying IPA-mediated neuroprotection. MethodsAn ICH model was established using C57BL/6 mice, which then received intragastric IPA (20 mg/kg/day). Fecal abundance of IPA-related genes and IPA levels in feces and plasma were measured by qPCR and UPLC-MS/MS. Behavioral tests, qPCR, and immunofluorescence staining were used to assess neurological function, myelin integrity, and axonal injury. In vitro, BV2 microglia, with or without Stap1 knockdown, were co-cultured with myelin debris to assess IPAs effects on phagocytosis. Additionally, targeted plasma IPA profiling was performed in 30 ICH patients and 30 matched healthy controls. ResultsThe relative abundance of IPA production-related genes and IPA levels in feces and plasma were significantly decreased after ICH and remained low into the chronical phase. After IPA administration, the neurological deficits, myelin loss and axonal injury of mice with ICH were significantly improved. In vitro, IPA increased BV2 microglia myelin debris phagocytosis by inhibiting Stap1 expression. IPA levels were significantly reduced in ICH patients, consistent with ICH mouse model. ConclusionsOur findings demonstrated that the gut microbiota-derived metabolite IPA could facilitate neurological deficits recovery and alleviate WMI, which could be a promising therapeutic strategy to improve ICH prognosis.

neuroscience↗

bioRxiv2025

Rice Ethylene Receptors Function as Ca2+-Permeable Ion Channels to Orchestrate Calcium-Dependent Antagonism of Ethylene Responses in Roots

The gaseous phytohormone ethylene governs essential plant processes spanning development, productivity, and stress resistance. As an essential nutrient and second messenger, calcium (Ca2+) is widely implicated in diverse plant physiological activities; however, its role in the ethylene signal transduction pathway remains elusive. Here, we identified a calcium-dependent antagonism of ethylene response (CAER) specifically modulates root elongation in the model cereal rice (Oryza sativa). Interestingly, we demonstrate that the ethylene receptors OsERS1 and OsERS2 both function as Ca2+-permeable channels, with OsERS1 further acting as a non-selective cation channel capable of permeating both monovalent and divalent cations. Mutagenesis analysis reveals that OsERS1 channel activity relies on homomeric assembly sites (C4/C6) rather than its ethylene-binding site (C65), indicating a clear decoupling of the molecular modules governing receptor signaling and ion channel function. The loss-of-function mutant Osers1/2 does not exhibit the CAER phenotype of the wild type (WT), confirm that this calcium-dependent regulatory mechanism is dependent on both OsERS1 and OsERS2. These findings uncover an unexpected ion-channel function of ethylene receptors, redefining their molecular identity beyond canonical signaling proteins and establishing the novel "hormone receptor-type ion channel (HRIC)" paradigm--one that fundamentally expands our understanding of how plant hormones transduce signals at the membrane.

developmental biology↗

bioRxiv2025

Diagnosing linearity along the carbon cascade in terrestrial biosphere models

Elevated carbon dioxide (eCO2) fertilises photosynthesis, driving an increase in terrestrial gross primary production (GPP). However, it is unclear how effectively increased GPP propagates along the "carbon (C) cascade" to increase net primary production (NPP) and vegetation C stocks (Cveg) in different plant compartments. Vegetation models simulate divergent C cycle projections and have been criticised for being overly photosynthesis-driven (source-driven), neglecting processes that lead to non-linear behaviour in response to the GPP increase, which may attenuate (or amplify) changes in NPP and vegetation C stocks. Here, we introduce an analytical framework to diagnose linearity (L) of the land C cycle as the ratio of relative changes in linked fluxes and pools and apply it to outputs from 16 models of the TRENDY v11 ensemble. We found widely varying patterns in L across models and for the different links. Six models showed a clear dominance of larger relative changes in NPP than in GPP in global simulations (LNPP:GPP >1 for >60% of gridcells), indicating increased carbon use efficiency under eCO2. Only three models had LNPP:GPP < 1 for >60% of gridcells. Four models showed a clear dominance of larger relative changes in steady-state Cveg than in NPP, while five models showed an opposite pattern - in both cases with a large spread of LCveg*:NPP across gridcells within models. Three models showed a larger relative increase in root C than in Cveg, while two models showed a clear dominance of the opposite pattern. Widely differing distributions of L across models and links reveal a strong influence of alternative process representations (nonlinear behaviour) in individual models. However, for all links, L deviations from 1 were roughly balanced across the model ensemble, leading to an overall linear behaviour of terrestrial C cycle representations.

ecology↗

bioRxiv2025

ALK1-BMPRII agonism by clustering bispecific antibodies treats hereditary hemorrhagic telangiectasia

Hereditary hemorrhagic telangiectasia (HHT) is characterized by arteriovenous malformations (AVMs) and severe bleeding caused by loss-of-function mutations in the ALK1 receptor pathway. We developed clustering agonist bispecific antibodies (BsAbs) targeting ALK1 and its activating partner, the Ser/Thr receptor kinase BMPRII. These BsAbs induced ALK1-BMPRII proximity association, stimulated the downstream Smad1/5/8 signaling cascade, and treated HHT pathologies in various mouse models. BsAb treatment reduced AVM burden by up to 95% in HHT mice, preventing anemia, cardiomegaly, and premature death. The BsAbs also enhanced Smad1/5/8 signaling in endothelial cells derived from HHT patients with ALK1 mutations and prevented retinal AVMs in a newly developed knock-in mouse carrying an HHT-causing ALK1 mutation. These findings establish ALK1-BMPRII agonism as a promising therapeutic strategy for HHT.

cell biology↗

bioRxiv2025

Deciphering the microbial landscape of lower respiratory tract in health and respiratory diseases

The microbiota plays an important role in maintaining lung homeostasis and the development of respiratory diseases. However, there have not been detailed characteristics of lung microbiome in health and respiratory diseases. Bronchoalveolar lavage fluid of 43 healthy individuals, 23 COVID patients, and 28 non-small cell lung cancer patients was conducted to characterize microbial diversity and function by metagenomic sequencing. We identified 196 species in health lung after removing low abnudance species. The most abundant species were Staphylococcus aureus, Salmonella enterica, and Klebsiella pneumoniae. Keystone species were identified through co-network analysis, such as Granulicatella adiacens and Mogibacterium diversum. To further explore the microbial function in the healthy lung, we obtained 56 non-redundant metagenome-assembled genomes (MAGs) and subsequently classified them into nine phyla. Metabolism and genetic information processing exhibit high abundance, including bacterial secretion systems, protein export, and Staphylococcus aureus infection, indicating a close correlation between microbiota and the host. Furthermore, the microbiome pairwise distances of non-small cell lung cancer (NSCLC) participants were more similar to those of the healthy group than those of individuals with COVID-19. Rothia dentocariosa, Corynebacterium kefirresidentii, Sphingobium yanoikuyae were enriched in the healthy group, while Schaalia odontolytica, Actinomyces graevenitzii, and Rothia mucilaginosa was enriched in the NSCLC group. Overall, this study provides a comprehensive overview of the diversity and gene function of the lung microbiome, which will facilitate future studies of microbiota associated with respiratory diseases in humans.

microbiology↗

bioRxiv2025

Tenotomized Achilles tendons of MRL/MpJ mice show scar-mediated healing, but have superior recovery of biomechanical properties

Tendon healing in Murphy Ross Large (MRL/MpJ) mice was examined using an Achilles tendon tenotomy model, a full transection model, compared to a patella focal injury model in which tendons in MRL/MpJ mice showed regenerative healing. After complete transection, the Achilles tendons of MRL/MpJ mice healed via scar formation, regardless of sex, as did C57BL/6J mice. Tensile testing found that mechanical properties of injured tendons of both MRL/MpJ female and male mice exhibited comparable levels to those of C57BL/6J female and male mice, respectively after 4 weeks of healing, which is during the remodeling phase. After 10 weeks of healing, injured tendons of MRL/MpJ mice possessed smaller heterotopic ossification volumes than those of C57BL/6J mice. Proteomics analysis revealed similar alterations to signaling pathways in injured tendons of MRL/MpJ and C57BL/6J male mice after 4 weeks of healing. However, among the altered pathways, actin cytoskeleton and integrin signaling pathways were two of the top pathways that were more prominently activated in C57BL/6J males than in MRL/MpJ males. These findings indicate that MRL/MpJ mice possess limited capacity to regenerate injured tendons after complete rupture, and that tenotomized Achilles tendons of MRL/MpJ male mice have lesser induction of heterotopic ossification and lower activation of the signaling pathways also induced with injury in C57BL/6J male mice.

pathology↗

bioRxiv2025

Unusually Broad-spectrum small-molecule sensing using a single protein scaffold

Small-molecule sensing in plants is dominated by chemical-induced dimerization modules. In the abscisic acid (ABA) system, allosteric receptors recruit phosphatase effectors and achieve nM in vivo responses from {micro}M receptor-ligand interactions. This sensitivity amplification could enable ABA receptors to serve as generic scaffolds for designing small-molecule sensors. To test this, we screened collections of mutant ABA-receptors against 2,726 drugs and other ligands and identified 569 sensors for 6.7% of these ligands. The mutational patterns indicate strong selection for ligand-specific binding pockets. We used these data to develop a sensor design pipeline and isolated sensors for multiple plant natural products, 2,4,6-trinitrotoluene (TNT), and "forever" per- and polyfluoroalkyl substances (PFAS). Thus, the ABA sensor system enables design and isolation of small-molecule sensors with broad chemical scope and antibody-like simplicity.

synthetic biology↗