Browse Articles
Discover research articles across all indexed journals
Template-free 3D programmable magnetization of soft millirobots induced by interlayer stress
Soft magnetic miniature devices are crucial for applications in minimally invasive medicine, soft electronics, and robotics. While substantial progress has been made, current magnetic programming techniques are inherently tied to template-based and sequential fabrication processes. These processes limit scalability, precision, and programmability. Here, we present a template-free, integrative strategy that leverages interlayer stress-induced 3D shape morphing in xerogel-PDMS bilayer materials triggered by temperature variations. This process induces preprogrammed deformation and fixes the 3D structure via interlayer stress and solid–liquid phase transition. It is akin to an insect encased in amber, resulting in a soft machine with precisely tailored magnetic domains upon saturated magnetization. The approach eliminates the need for predesigned molds, which offers scalable, template-free programmable magnetization, reducing time and labor costs. The versatility of this method is demonstrated through reconfigurable mechanical behavior in kirigami metamaterial structures, information encryption, and multilegged millirobots. Moreover, by incorporating a nonmagnetic PDMS layer, laser-based engraving and ablation allow simultaneous control of interlayer stress and material properties. This facilitates precise regulation of stress-induced deformation and magnetically responsive regions with 20 μm resolution and over 1.8 T magnetization strength. This template-free 3D magnetization strategy significantly enhances design flexibility, machining precision, and mass production. It paves the way for advanced multiscale and programmable soft magnetic devices.
Stepwise deactivation of gibberellins during rice internode elongation
Bioactive gibberellins (GAs) are a class of plant hormones that regulate various aspects of plant growth and development, and several key GA deactivation enzymes have been identified. In rice, non-13-hydroxylated GAs have been shown to be deactivated via 16α,17-epoxidation by a cytochrome P450 monooxygenase, ELONGATED UPPERMOST INTERNODE (EUI/CYP714D1). Although 16,17-dihydro-16α,17-epoxyGA 4 (16α,17-epoxyGA 4 ), the product of EUI from bioactive GA 4 , has shown weak bioactivity on rice seedlings, how 16α,17-epoxyGAs are further deactivated remains elusive. Here, we identify the EUI2 gene, which regulates internode elongation in rice, using a map-based cloning strategy. EUI2 encodes an epoxide hydrolase that hydrolyzes 16α,17-epoxyGAs to 16,17-dihydro-16α,17-dihydroxyGAs. The eui2 mutants are taller than wild-type plants but are shorter than the eui mutants. However, the levels of known bioactive GAs in the uppermost internodes are not significantly increased in the eui2 mutants. Instead, we show that the eui2 mutants accumulate 16α,17-epoxyGA 4 to high levels. We also show that exogenously applied 16α,17-epoxyGA 4 is significantly active in elongating the uppermost internode, although not as potent as GA 4 . Furthermore, we demonstrate that 16α,17-epoxyGA 4 can directly interact with the rice GA receptor, GIBBERELLIN INSENSITIVE DWARF1, in vitro. Taken together, the sequential action of EUI and EUI2 results in the stepwise deactivation of GAs during internode elongation in rice. Our data also suggest that the accumulation of a weakly active GA contributes to the mildly tall phenotype of the eui2 mutants.
Direct sensing of host ferric iron by an archetype histidine kinase mediates virulence of an enteric pathogen
Two-component system (TCS) histidine kinases enable bacterial pathogens to sense environmental signals and regulate adaptive responses during infection. The EnvZ/OmpR TCS, known for its role in osmolarity/pH-dependent regulation of outer membrane porins across bacterial species, is also a central virulence regulator. However, the environmental cues that activate EnvZ/OmpR to trigger pathogenicity have remained unclear, limiting our understanding of host–pathogen interactions. Here, we demonstrate that in Vibrio parahaemolyticus , a major etiological agent of seafood-associated gastroenteritis, EnvZ functions as a direct ferric iron (Fe 3+ ) sensor governing virulence programs. Fe 3+ -EnvZ interaction triggers kinase phosphorylation and activation, enabling transcriptional control of biofilm formation, swarming motility, and type 3/6 secretion systems. An iron-binding-deficient EnvZ mutant (EnvZ Q103A ) abrogated Fe 3+ responsiveness and downstream signaling pathways. In an infant rabbit infection model, Fe 3+ enhanced V. parahaemolyticus intestinal colonization and virulence through EnvZ/OmpR signaling. This study identifies Fe 3+ as the physiological ligand activating the EnvZ/OmpR virulence regulon and provides insight into how enteric pathogens exploit host-derived iron cues to promote infection.
Multiphasic size-dependent growth dynamics of nanoparticle ensembles
Colloidal nanoparticles are of great interest in modern science and industry. However, the thermodynamic mechanism and dynamics of nanoparticle growth have yet to be understood. Addressing these issues, we tracked hundreds of in-situ growth trajectories of a nanoparticle ensemble using liquid-phase TEM and found that the nanoparticle growth, including coalescence, exhibits nanoparticle size-dependent multiphasic dynamics, unexplainable by current theories. Motivated by this finding, we developed a model and theory for an ensemble of growing nanoparticles, providing a unified, quantitative understanding of the time-dependent mean and fluctuation of nanoparticle size and size-dependent growth rate profiles across various nanoparticle systems and experimental conditions. Our work reveals that the chemical potential in a small nanoparticle strongly deviates from the Gibbs–Thomson equation, shedding light on how it governs the size-dependent growth dynamics of nanoparticles.
The analgesic paracetamol metabolite AM404 acts peripherally to directly inhibit sodium channels
Paracetamol has been used for decades to relieve mild-to-moderate pain. Its analgesic effect is mainly attributed to its metabolite, AM404, acting on cannabinoid receptors or TRPV1 channels in central nervous system (CNS) neurons. Here, we show that AM404 is produced by primary sensory neurons. It inhibits sodium current in nociceptor neurons, blocking action potential (AP) generation and reducing nocifensive behavior in naïve and inflamed rats. We demonstrated that this analgesic effect of AM404 is mediated by its direct inhibition of nociceptive voltage-gated sodium channels (Na V ) 1.8 and 1.7 via the local anesthetic binding site. The Na V 1.8 and 1.7 inhibition was specific for AM404 and not observed with other metabolites of paracetamol. Our findings suggest that the analgesic effect of paracetamol is mediated mainly by direct AM404-induced inhibition of nociceptive sodium channels at the peripheral nociceptor neurons. Our findings lay a foundation for the potential development of AM404 as a selective local analgesic.
Spatially resolved DNP-assisted NMR illuminates the conformational ensemble of α-synuclein in intact viable cells
The protein α-syn adopts a wide variety of conformations including an intrinsically disordered monomeric form and an α-helical-rich membrane-associated form that is thought to play an important role in cellular membrane processes. However, despite the high affinity of α-syn for membranes, evidence that the α-helical form is adopted inside cells has been indirect. DNP-assisted solid-state NMR on frozen cellular samples can report directly on the entire conformational ensemble. By controlling the distribution of the DNP agent throughout the cellular biomass, such experiments can provide quantitative information upon the entire structural ensemble or provide information about spatially resolved subpopulations. When the polarization agent is dispersed homogeneously throughout the cell, a minority of the α-syn inside HEK293 cells adopts a highly α-helical-rich conformation. When the polarization agent is peripherally localized, the α-helical-rich conformation predominates, indicating that it is preferentially adopted near the cellular periphery. This demonstrates how selectively altering the spatial distribution of the DNP agent can be a powerful tool to observe spatially distinct structural ensembles. This approach paves the way for more nuanced investigations into the conformations that proteins adopt in different areas of the cell.
Transcranial direct current stimulation neuromodulates intracranial cognitive evoked activity in humans
Transcranial direct current stimulation (tDCS) is an easy to use, noninvasive brain stimulation technique that gained prominence for its potential in cognitive rehabilitation. Electroencephalography (EEG), which records electrical brain activity with a high temporal resolution, is well suited to quantify tDCS-induced neuromodulation in humans. However, most studies relying on scalp EEG recordings or event-related potentials showed low reliability and only indirect correlations. Here, we combined intracranial EEG (iEEG) recordings with a sham-controlled tDCS experiment during fast periodic visual stimulation. Anodal (+2 mA) tDCS was applied over the right occipito-temporal cortex for 20 min using two small ring high-definition electrodes. Through the analysis of iEEG signals of 947 intracerebral contacts in 11 drug-resistant epileptic patients, we quantified the neuromodulation of iEEG cognitive evoked responses during (P2 phase) and after (P3 phase) tDCS by comparison to a control phase before tDCS (P1 phase). Significant neuromodulations of face-selective iEEG activity in anterior & posterior temporal lobe and in the occipital lobe were found, with amplitude increases of 3% and 4%, 16% and 13%, and 36% and 33%, during and after tDCS, respectively. Interestingly, despite a unique tDCS session, the face-selective neuromodulation in the right visual occipito-temporal cortex remained significant ( P = 0.015) after tDCS (P3 vs. P1). This iEEG study demonstrates that using low intensity tDCS and small ring electrodes can induce significant electrophysiological effects on a selective cognitive function in humans.
High-throughput metabolic engineering of <i>Yarrowia lipolytica</i> through gene expression tuning
The challenge of accurately predicting which genetic alternations lead to the desired phenotype necessitates high-throughput metabolic engineering approaches where numerous hypotheses can be tested simultaneously. We describe the CRISPR-Cas9-based method TUNE YALI that enables high-throughput tuning of gene expression in the common industrial yeast Yarrowia lipolytica . The method is based on replacing the promoters of the target genes with native Y. lipolytica promoters of varying strengths or removing the promoters entirely. To demonstrate the method’s capabilities, we created a plasmid library that targets 56 transcription factors (TFs) and changes the expression of each TF to seven different levels. We transformed this library into reference and betanin-producing strains of Y. lipolytica and screened the resulting clones for changes in morphology, thermotolerance, or improved betanin production. The genetic markup of the yeast clones with the desired phenotypic changes was determined by sequencing the inserted plasmids. We identified multiple TFs whose regulatory changes increased thermotolerance, two TFs that eliminated pseudohyphal growth, and several TFs that increased betanin production. Analogous libraries can be designed to target any chosen group of genes and even all the genes. The libraries can be shared and reused, accelerating applied strain development projects and fundamental functional genomics research (TUNE YALI -TF kit and TUNE YALI -TF library are available via AddGene under catalog numbers #1000000255 and #217744).
Parallel sensory compensation following independent subterranean colonization by groundwater salamanders ( <i>Eurycea</i> )
Lineages that have invaded subterranean environments have repeatedly evolved remarkable adaptations to life in darkness. However, observational and experimental studies in additional natural systems are needed to further our understanding of repeated evolution and convergence. In Texas, a radiation of groundwater salamanders (genus Eurycea ), with independent invasions of subterranean karstic environments, offers an opportunity to investigate phenotypic convergence, parallel evolution, and the enhancement and regression of sensory systems. Adaptations to a troglobitic life in this clade include morphological, behavioral, and physiological changes within and among species. Intraspecific and interspecific variation in morphology in response to the selective pressures of life underground allows for detailed examination of physical, behavioral, and physiological changes associated with subterranean adaptation within a comparative phylogenetic framework. We find a correlated change between two sensory systems repeated across multiple subterranean Eurycea lineages: the degeneration of the eye and the expansion of the mechanosensory lateral line. The increase in anterior neuromast organs in subterranean lineages was positively correlated with the expression of pax6 (Paired-box 6), a conserved transcription factor important for vertebrate neurogenesis. Our results show a decreasing trend of PAX6 labeling in the neuromasts of adult surface salamanders ( Eurycea nana ) relative to the maintained labeling in subterranean species ( Eurycea rathbuni ). These lateral line enhancements are correlated with reductions in the development of optic systems in subterranean salamander lineages. Altogether, our findings provide a starting point for future evolutionary developmental investigations examining the genetic underpinnings of adaptive, repeated evolution in a novel system.
Polaron catastrophe within quantum acoustics
The quantum acoustic framework has recently emerged as a nonperturbative, coherent approach to electron–lattice interactions, uncovering rich physics often obscured by perturbative methods with incoherent scattering events. Here, we model the strongly coupled dynamics of electrons and acoustic lattice vibrations within this framework, representing lattice vibrations as coherent states and electrons as quantum wave packets, in a manner distinctively different from tight-binding or discrete hopping-based approaches. We derive and numerically implement electron backaction on the lattice, providing both visual and quantitative insights into electron wave packet evolution and the formation of acoustic polarons. We investigate polaron binding energies across varying material parameters and compute key observables—including mean square displacement, kinetic energy, potential energy, and vibrational energy—over time. Our findings reveal the conditions that favor polaron formation, which is enhanced by low temperatures, high deformation potential constants, slow sound velocities, and high effective masses. Additionally, we explore the impact of external electric and magnetic fields, showing that while polaron formation remains robust under moderate fields, it is weakly suppressed at higher field strengths. These results deepen our understanding of polaron dynamics and pave the way for future studies into nontrivial transport behavior in quantum materials.
The prevalence of functional limitations in the US workforce
This research paper investigates the prevalence of functional limitations among employed adults in the United States and the association between these limitations and medical conditions. The authors administered a survey adapted from the Dutch Functional Abilities List to a nationally representative sample of US adults ages 22 and older, finding that nearly three-quarters of working adults report at least one functional limitation, with an average of nearly six functional limitations per working adult. The most common limitations were in upper body strength and torso range of motion, and with respect to the ambient environment. The study also found that mental illness, arthritis, and substance use disorder are associated with the greatest number of functional limitations in working adults. The findings have implications for economic performance, workforce planning, and social policies to support displaced and vulnerable workers with significant functional limitations. Identifying the occupations and industries with large numbers of workers with functional limitations is critical to addressing short-term labor supply disruptions (e.g., public health crises) and preparing for longer-term workforce needs (e.g., long-term care workers for an aging population).
Detection of the knee point in lithium-ion battery degradation using a state-of-charge-dependent parameter
The rapidly expanding lithium-ion battery (LIB) market has heightened the demand for efficient diagnostics for in-use cells and the reliable grading of used cells. Various purpose-built analysis tools and statistical algorithms have been developed, but often rely on redundant instrumentation and computationally intensive procedures. Here, we propose using the variance of the capacity difference between 0.2C and 1C, Var(Δ Q 0.2C-1C (V) ), based on the strong correlation between the interfacial state of the anode and mode of capacity degradation, as a measure of the health state of individual cells. A single-point “off-board” measurement of Var(Δ Q 0.2C-1C (V) ) indicates whether a particular cell is experiencing self-limiting or accelerating degradation and is thus near a knee point in its cycle life. This assessment additionally provides a quantitative criterion for differentiating used cells for reuse or recycling. Our findings suggest that utilizing state-of-charge-dependent key electrochemical properties enables the cell health to be accurately monitored, thereby promoting sustainability in the expanding battery market.
Natural dispersal is better than translocation for reducing risks of inbreeding depression in eastern black rhinoceros ( <i>Diceros bicornis michaeli</i> )
Due to increasing anthropogenic impacts, many species survive only in small and isolated populations. Active conservation management to reduce extinction risk includes increasing habitat connectivity, translocations from captive populations, or intensive surveillance of highly protected closed populations. Advances in sequencing technology mean that it is now possible to consider the genomic impacts of such strategies, as a proxy for variation in individual fitness. Using whole genome sequences from critically endangered eastern black rhinoceros ( Diceros bicornis michaeli ), we compare the consequences of different types of conservation efforts, based on cohorts of offspring resulting from parents from different sources. Based on the fraction of the genome in runs of homozygosity (ROH) of different lengths, we found lower inbreeding in offspring of individuals that had either been translocated from ex-situ populations (F ROH>1Mb = 0.047) or dispersed between proximate native populations (F ROH>1Mb = 0.065) compared to the intensively managed closed population from which the migrant moved (F ROH>1Mb = 0.112). However, the benefit of such movement was removed after only a few generations of closed breeding (F ROH>1Mb = 0.149). Although sample size restricted power to detect significance of differences, the relative abundance of highly deleterious mutations was higher for offspring resulting from translocation compared to the other cohorts and this load was sheltered by higher heterozygosity, which could increase risks of inbreeding depression if inbreeding subsequently occurs. In contrast, native dispersers reduced the negative effects of inbreeding without compromising the benefits of past purging of deleterious mutations. Our study highlights the importance of natural dispersal and reiterates the importance of maintaining habitat corridors between populations.
Epithelial Regnase-1 inhibits colorectal tumor growth by regulating IL-17 signaling via degradation of <i>NFKBIZ</i> mRNA
Regnase-1 is a ribonuclease that regulates inflammation in immune cells by degrading cytokine mRNA. Regnase-1 was identified as one of the frequently mutated genes in the inflamed colorectal epithelium of patients with ulcerative colitis; however, its significance in intestinal epithelial cells during the tumorigenic process remains unknown. Therefore, we developed an Apc Min/+ mouse model lacking Regnase-1 in intestinal epithelia. Regnase-1 deletion significantly enhanced colon tumor growth accompanied by elevated levels of extracellular signal-regulated kinase (ERK) phosphorylation in tumor tissues. Transcriptome analysis of the tumor tissues revealed that Nfkbiz , a mediator of the interleukin (IL)-17 signaling pathway, was the primary degradative target of Regnase-1 in enterocytes and that Regnase-1 deficiency enhanced IL-17 signaling. The treatment with antibiotics or IL-17-neutralizing antibody canceled the proliferative effect of colon tumors due to Regnase-1 deletion, suggesting the protective role of Regnase-1 against colon tumor growth was dependent on IL-17 signaling triggered by gut microbes. Analysis of the Nfkbiz knockout mouse model demonstrated that the tumor-suppressive effect of Regnase-1 depended on Nfkbiz expression. Remarkably, oral treatment of dimethyl fumarate, a potential inhibitor of Regnase-1 protein inactivation, suppressed tumor growth, downregulated Nfkbiz , and suppressed ERK activation. Furthermore, TCGA data analysis revealed that low Regnase-1 expression in colorectal cancer tissue was related to poor prognosis. Therefore, Regnase-1 represses colon tumor growth by regulating IL-17 signaling via Nfkbiz mRNA degradation. Regnase-1 could be a potential therapeutic target in colon tumors.
Study design and the sampling of deleterious rare variants in biobank-scale datasets
One key component of study design in population genetics is the “geographic breadth” of a sample (i.e., how broad a region across which individuals are sampled). How the geographic breadth of a sample impacts observations of rare, deleterious variants is unclear, even though such variants are of particular interest for biomedical and evolutionary applications. Here, in order to gain insight into the effects of sample design on ascertained genetic variants, we formulate a stochastic model of dispersal, genetic drift, selection, mutation, and geographically concentrated sampling. We use this model to understand the effects of the geographic breadth of sampling effort on the discovery of negatively selected variants. We find that samples which are more geographically broad will discover a greater number of variants as compared to geographically narrow samples (an effect we label “discovery”); though the variants will be detected at lower average frequency than in narrow samples (e.g., as singletons, an effect we label “dilution”). Importantly, these effects are amplified for larger sample sizes and fitness effects. We validate these results using both population genetic simulations and empirical analyses in the UK Biobank. Our results are particularly important in two contexts: the association of large-effect rare variants with particular phenotypes and the inference of negative selection from allele frequency data. Overall, our findings emphasize the importance of considering geographic breadth when designing and carrying out genetic studies, especially at biobank scale.
Population sequencing for phylogenetic diversity and transmission analyses
Genomic diversity in pathogen populations is foundational for evolution and adaptation. Understanding population-level diversity is also essential for tracking sources and revealing detailed pathways of transmission and spread. For bacteria, culturing, isolating, and sequencing the large number of individual colonies required to adequately sample diversity can be prohibitively time-consuming and expensive. While sequencing directly from a mixed population will show variants among reads, they cannot be linked to reveal allele combinations associated with phylogenetic inheritance patterns. Here, we describe the theory and method for using population sequencing directly from a mixed sample, along with a minimal number of individually sequenced colonies, to describe the phylogenetic diversity of a population without haplotype reconstruction. To demonstrate the utility of population sequencing in capturing phylogenetic diversity, we compared isogenic clones to population sequences of Burkholderia pseudomallei from sputum of a single patient. Our results point to the pathogen population being highly structured, suggesting that for some pathogens, sputum sampling may preserve structuring in the lungs and thus present a noninvasive alternative to understanding colonization, movement, and pathogen/host interactions. We also analyzed population sequences of Staphylococcus aureus derived from different people and different body sites to reveal directionality of transmission between hosts and across body sites, demonstrating the power and utility for characterizing the spread of disease and identification of reservoirs at the finest levels. We anticipate that population sequencing and analysis can be broadly applied to accelerate research in a wide range of fields reliant on a foundational understanding of population phylogenetic diversity.
Deciphering decomposition pathways of high explosives with cryogenic X-ray Raman spectroscopy
We employed cryogenic X-ray Raman spectroscopy to investigate the early-stage decomposition of the high explosive molecule hexanitrohexaazaisowurtzitane (CL-20). By systematically varying the radiation dose under cryogenic conditions, we induced the decomposition of the molecule using ionizing radiation and observed the evolution of spectral features at the carbon, nitrogen, and oxygen K edges. Through extensive first-principles calculations, we identified key intermediates in the early stages of the decomposition process, resulting from C–C and C–N bond cleavage which leads to the opening of the internal cage structure. A detailed analysis of spectral trends and fingerprints provided evidence supporting N–NO 2 homolytic cleavage as the primary initial decomposition pathway. The combination of advanced core-level spectroscopy methods and state-of-the-art theoretical calculations enabled a comprehensive characterization of the molecular changes induced by controlled radiation dose exposures. Our findings establish a benchmark for understanding the decomposition chemistry of high-explosive materials, offering important insights into their stability and reactivity under extreme conditions.
<i>CACNA1D</i> is a circadian gene and causes familial advanced sleep phase
Familial advanced sleep phase (FASP) is a heritable human sleep trait characterized by early sleep onset and offset times. We have identified five variants in five different families in the human voltage-gated calcium channel subunit alpha1 D ( CACNA1D ) that cosegregate with FASP. The variants in CACNA1D lead to altered channel dynamics in vitro. A mouse model of the E427K variant has a normal circadian period under constant darkness but displays altered phase shifts in response to light in the subjective night at circadian time (CT) 16 and CT22. Overall, these experiments establish CACNA1D as an FASP gene with altered entrainment, highlighting the ability of human genetics to uncover novel aspects of human circadian regulation.
BRCA2 reversion mutation–independent resistance to PARP inhibition through impaired DNA prereplication complex function
Recent approvals of polymeric adenosine diphosphate ribose (poly(ADP-ribose) polymerase inhibitors (PARPi) for BRCA-mutant metastatic castration resistant prostate cancer necessitate an understanding of the factors that shape sensitivity and resistance. Reversion mutations that restore homologous recombination (HR) repair are detected in ~50 to 80% of BRCA-mutant patients who respond but subsequently relapse, but there is currently little insight into why only ~50% of BRCA-mutant patients display upfront resistance. To address this question, we performed a genome-wide CRISPR screen to identify genomic determinants of PARPi resistance in murine Brca2 Δ/Δ prostate organoids genetically engineered in a manner that precludes the development of reversion mutations. Remarkably, we recovered multiple independent single guide RNAs (sgRNAs) targeting three different members ( Cdt1, Cdc6, and Dbf4 ) of the DNA prereplication complex (pre-RC), each of which independently conferred resistance to olaparib and the next-generation PARP-1 selective inhibitor AZD5305. Moreover, sensitivity to PARP inhibition was restored in Brca2 Δ/Δ , Cdc6-depleted prostate cells by knockdown of geminin, a negative regulator of Cdt1, further implicating the critical role of a functional pre-RC complex in PARPi sensitivity. Furthermore, ~50% of CRPC tumors have copy number loss of pre-RC complex genes, particularly CDT1 . Mechanistically, prostate cells with impaired pre-RC activity displayed rapid resolution of olaparib-induced DNA damage as well as protection from replication fork degradation caused by Brca2 loss, providing insight into how Brca2-mutant cancer cells can escape cell death from replication stress induced by PARP inhibition in the absence of HR repair. Of note, a pharmacologic inhibitor that targets the CDT1/geminin complex (AF615) restored sensitivity to AZD5305, providing a potential translational avenue to enhance sensitivity to PARP inhibition.
Disrupted diencephalon development and neuropeptidergic pathways in zebrafish with autism-risk mutations
Hundreds of human mutations are linked to autism and related disorders, yet the functions of many of these mutated genes during vertebrate neurodevelopment are unclear. We generated 27 zebrafish mutants with presumptive protein-truncating mutations or specific missense variants corresponding to autism-risk alleles in 17 human genes. We observed baseline and stimulus-driven behavioral changes at larval stages, as well as social behavior differences in lines tested as juveniles. Imaging whole-brain activity revealed a near identical activity map for mutations in the unrelated genes kmt5b and hdlbpa , defined by increased activity mainly in the thalamus and mesencephalon. Mutating 7 of the 17 risk genes resulted in substantial brain size differences, localized to the diencephalon in three cases and more widespread in others. Using RNA sequencing, we further defined molecular drivers of the observed phenotypes for three mutants, identifying targetable disruptions in neuropeptide signaling, neuronal maturation, and cell proliferation. This multimodal screen nominated brain regions, cell types, and molecular pathways that may contribute to autism susceptibility.