The recent revolution in genome sequencing and protein structure prediction has opened new frontiers in understanding, predicting and designing enzyme function. Central to these efforts is the discovery and functional an...The recent revolution in genome sequencing and protein structure prediction has opened new frontiers in understanding, predicting and designing enzyme function. Central to these efforts is the discovery and functional annotation of novel enzymes, which is essential for elucidating the connection between genotype and phenotype and for developing biocatalysts for industrial applications. However, accurately predicting enzymatic function remains a major challenge, and the discovery of new enzymes often relies on serendipity. Here we present a metal-coordination-guided strategy that uses atomic-level mechanistic principles to mine protein structure databases for the targeted discovery of metalloenzymes. We apply this framework to the AlphaFold2 Protein Structure Database to identify new members of the Fe/α-ketoglutarate-dependent halogenase family, which selectively functionalize unactivated C(sp)-H-bonds, a crucial transformation in the production of pharmaceuticals and other high-value compounds. These radical halogenases constitute a low-abundance class within the large and diverse cupin superfamily. Owing to low sequence conservation, they have been especially challenging to find against the complex background of related family members, such as hydroxylases, desaturases and epimerases. Our metal-coordination mining methodology reveals several previously unrecognized radical halogenase families spanning diverse phylogenetic space, at minimal computational cost. Our predictions are validated by the experimental characterization of two new radical halogenases, AspX and BtnX. Notably, BtnX shows a substrate promiscuity that is unprecedented in radical halogenases, opening the way for a broad range of biocatalytic applications.
Cui T, Wang S, Lee SS
… +17 more, Barks E, Cattermull J, Melamed C, Jiang Z, Morrison M, Narun L, Tzeng YK, Fujii N, Kim SH, Xu X, McConohy G, Wallace PM, Lee AC, Cui X, Lee JH, Chueh WC, Gu XW
Lithium-metal solid-state batteries offer advantages of high energy density and improved safety compared with lithium-ion batteries. However, solid-state batteries fail through short-circuiting even at low charging rates...Lithium-metal solid-state batteries offer advantages of high energy density and improved safety compared with lithium-ion batteries. However, solid-state batteries fail through short-circuiting even at low charging rates (less than 1 mA cm) due to lithium dendrite initiation and propagation. The location of dendrite initiation is under debate, particularly regarding whether initiation occurs within the interior of the solid electrolyte or at the surface. Here we develop an in-plane biaxial compression method that provides direct evidence that dendrite initiation occurs within the interior of garnet LiLaZrTaO solid electrolytes during long-term cycling when the surface initiation mechanisms are rendered ineffective in shorting the cell. The biaxial compression deflects dendrite propagation so that it is perpendicular to the electric field direction, leading to the generation of an unprecedentedly high density of dendrites without short-circuiting, even at an extreme fast-charging rate of 100 mA cm. After long-term cycling, dendrites eventually appeared throughout the entire thickness of the solid electrolyte. Under extreme cycling conditions, isolated lithium deposits are observed at grain-boundary junctions and pores, and these act as the dendrite initiation sites. This work reconciles the surface and interior initiation mechanisms in garnet solid electrolytes and demonstrates that in-plane biaxial compressive stress can prevent both from short-circuiting the cell.
Voluntary muscle contraction is triggered by the neurotransmitter acetylcholine binding its receptors on the postsynaptic membrane of the neuromuscular junction, opening ion channels that allow cation influx and initiate...Voluntary muscle contraction is triggered by the neurotransmitter acetylcholine binding its receptors on the postsynaptic membrane of the neuromuscular junction, opening ion channels that allow cation influx and initiate depolarization. Mutations in muscle acetylcholine receptors disrupt this process by either impairing (fast-channel) or prolonging (slow-channel) channel openings. These defects cause congenital myasthenic syndromes (CMS), characterized by severe muscle weakness that is often present at birth and, in some cases, progresses to paralysis and death. The structural mechanisms underlying these pathogenic defects and their pharmacological correction remain unknown. Here, using cryogenic electron microscopy, chemical biology and electrophysiology, we determined the structures and functional consequences of representative CMS mutant receptors with and without drugs. In fast-channel disease-associated mutants, we discovered a cryptic allosteric site targeted by positive modulators that restore gating in a mutation-specific manner. In receptor mutants associated with slow-channel disease, quinidine, fluoxetine and reboxetine act as pore blockers; notably, the antidepressant reboxetine selectively blocks desensitized receptors in a mutation-independent fashion, suggesting repurposing potential. Mechanistically, fast-channel mutations uncouple agonist binding from gating, whereas slow-channel mutations stabilize an abnormally widened, desensitized-like pore. These findings reveal unifying principles of CMS pathogenesis and provide a framework for precision therapies.
Sorensen SA, Gouwens NW, Wang Y
… +115 more, Mallory M, Budzillo A, Dalley R, Lee BR, Gliko O, Kuo HC, Kuang X, Mann R, Ahmadinia L, Alfiler L, Baftizadeh F, Baker KS, Bannick S, Bertagnolli D, Bickley K, Bohn P, Bomben J, Bowman C, Boyer G, Brouner K, Brown D, Cahoon A, Chen N, Chen C, Chen K, Chvilicek M, Collman F, Daigle TL, Dawes T, de Frates R, Dee N, DePartee M, Egdorf T, El-Hifnawi L, Enstrom R, Esposito L, Farrell C, Gala R, Gamlin C, Gary A, Glomb A, Gerasymchuk O, Goldy J, Gu H, Hadley K, Hawrylycz M, Henry A, Hill D, Hirokawa KE, Huang Z, Johnson K, Juneau Z, Kebede S, Kim L, Kruse L, Lee C, Leon AL, Lesnar P, Lheureux Q, Li A, Li Y, Liang E, Link K, Maxwell M, McGraw M, McMillen DA, Mukora A, Ng L, Ochoa T, Oldre A, Park D, Pom CA, Popovich Z, Potekhina L, Rajanbabu R, Ransford S, Reding MR, Ruiz A, Sandman D, Schroedter M, Sevigny J, Shulga L, Siverts L, Slaughterbeck CR, Smith KA, Stoecklin M, Sulc J, Sunkin SM, Tieu M, Ting JT, Trinh J, Velasco R, Vargas S, Vumbaco D, Walker M, Wang M, Wanner A, Waters J, Wells M, Williams G, Wilson JA, Xiong W, Lein ES, Berg J, Kalmbach BE, Yao S, Gong H, Luo Q, Wang Q, Ng L, Sümbül U, Yao Z, Jarsky T, Tasic B, Zeng H
The mammalian brain consists of diverse neuron types with various functions. Recent single-cell RNA sequencing approaches have led to a whole-brain taxonomy of transcriptomically defined cell types. Patch-seq experiments...The mammalian brain consists of diverse neuron types with various functions. Recent single-cell RNA sequencing approaches have led to a whole-brain taxonomy of transcriptomically defined cell types. Patch-seq experiments augment these cell-type descriptions by linking transcriptomic profiles with local morphological and electrophysiological properties. However, linking transcriptomic identities to long-range axonal projections remains a major unresolved challenge. Here, to address this, we collected two datasets from the mouse visual cortex consisting of: (1) 1,528 excitatory Patch-seq neurons, with local morphological, electrophysiological and transcriptomic data collected from each cell, and (2) 341 excitatory, whole-neuron morphologies. From the Patch-seq data, we defined 17 morphoelectric-transcriptomic types and built a multistep classifier to integrate cell-type assignments with whole-neuron morphology and interrogate cross-modality relationships. We find that transcriptomic variation within and across morphoelectric-transcriptomic types corresponds with morphological and electrophysiological phenotypes. In addition, these gene expression patterns, along with the anatomical location of the cell, can be used to predict projection targets of individual neurons. We observed novel multimodal cell-type signatures for layer 5 intratelencephalic and extratelencephalic neurons and shed new light on their axonal circuitry, including interhemispheric intratelencephalic projections. With this approach, we establish a comprehensive, integrated taxonomy of cortical, excitatory neuron types, and create a system for high-dimensional cell-type classification that can be extended to the whole brain and potentially across species.
Schiffers S, Nelson BW, Prigge M
… +12 more, Krishna S, Watkins L, Zhu Y, Tyagi N, Beiki H, Das S, Raman A, Ma J, Andresson T, Mao HQ, Wu B, Oberdoerffer S
Synthetic mRNA therapeutics offer a versatile platform for treating diverse conditions, including cancer and infectious diseases. For delivery into cells, these mRNAs are encapsulated in lipid nanoparticles and commonly...Synthetic mRNA therapeutics offer a versatile platform for treating diverse conditions, including cancer and infectious diseases. For delivery into cells, these mRNAs are encapsulated in lipid nanoparticles and commonly incorporate modified ribonucleotides to improve stability, enhance translation and mitigate immune recognition. N-Methylpseudouridine (mΨ) has become the industry standard for synthetic mRNAs owing to its effectiveness in promoting translation and reducing immunogenicity. However, recent studies have shown that mΨ can compromise translational fidelity, leading to errors such as premature termination and ribosomal frameshifting. Here we reveal N-acetylcytidine (acC) as a functionally distinct alternative to mΨ. Across cultured cell lines, primary human monocyte-derived dendritic cells and mouse liver, acC suppressed inflammatory responses as effectively as mΨ while driving higher protein yields. Single-molecule imaging of translation revealed broadly similar ribosome densities per mRNA for acC-modified and mΨ-modified transcripts. However, translation elongation with mΨ-modified mRNA was nearly twofold slower than with acC, which resulted in reduced protein output and increased ribosome collisions that further limited protein production through the engagement of quality-control pathways and +1 frameshifting. These findings underscore the importance of context in designing therapeutic mRNAs and position the translation elongation rate as a key determinant of the efficacy of modified ribonucleotides.
Huntington's disease (HD) is a devastating movement disorder without a cure at present. Although the monogenic basis of HD is well defined, the complex downstream effects that underlie behavioural symptoms are poorly und...Huntington's disease (HD) is a devastating movement disorder without a cure at present. Although the monogenic basis of HD is well defined, the complex downstream effects that underlie behavioural symptoms are poorly understood. These effects include cortical dysfunction, yet the roles of specific cortical neuronal subtypes in HD symptoms remain largely unexplored. Here we used longitudinal in vivo two-photon calcium imaging to examine the activity of three cortical inhibitory neuron (IN) subtypes and excitatory corticostriatal (CStr) projection neurons in the motor cortex of the transgenic R6/2 HD mouse model throughout disease progression. We found that motor deficits in R6/2 mice were accompanied by neuron subtype-specific abnormalities in movement-related activity. This included marked hypoactivity of vasoactive intestinal peptide (VIP)-INs and CStr neurons, which was also observed in the knock-in zQ175DN HD mouse model. Optogenetic activation of VIP-INs in R6/2 mice restored healthy levels of activity in VIP-INs and their downstream CStr neurons and ameliorated motor deficits in R6/2 mice; behavioural improvements persisted for days after stimulation. Our findings highlight cortical INs as a potential therapeutic target for HD.
Tin (Sn) halide perovskites are promising lead-free semiconductors for optoelectronic and electronic devices, owing to their tunable bandgaps and favourable charge transport. However, their practical implementation is fu...Tin (Sn) halide perovskites are promising lead-free semiconductors for optoelectronic and electronic devices, owing to their tunable bandgaps and favourable charge transport. However, their practical implementation is fundamentally limited by an intrinsic redox instability at undercoordinated Sn sites, which drives uncontrolled self-p-doping and rapid oxidative degradation. Here we introduce a volatile-assisted coordination strategy that reconstructs the perovskite surface through transient acetate coordination and volatilization, which transforms reactive SnI-terminated surfaces into chemically equilibrated and defect-mitigated interfaces. This surface reconstruction suppresses undercoordinated Sn-related trap states and stabilizes the local stoichiometry, thus enabling p-type transistors with robust transport characteristics, a near-zero threshold voltage and high on/off ratios exceeding 10. More importantly, the reconstructed interface acts as a self-passivating and thermally resilient barrier, resulting in markedly enhanced environmental stability, with devices maintaining stable operation for over 1 month at 100 °C. These results establish volatile-assisted surface reconstruction as an effective method for defect equilibration in metastable semiconductors, and they provide a general strategy for enabling durable, device-grade functionality in Sn-based materials.
Stratospheric water vapour (SWV) is a key greenhouse gas that influences both global climate and stratospheric ozone chemistry. Its abundance is strongly modulated by natural climate variability. Volcanic eruptions have...Stratospheric water vapour (SWV) is a key greenhouse gas that influences both global climate and stratospheric ozone chemistry. Its abundance is strongly modulated by natural climate variability. Volcanic eruptions have long been expected to humidify the stratosphere via tropopause warming, but observational confirmation has been lacking. Here we provide observational evidence that moderate volcanic eruptions and extreme wildfires since 2005 have systematically increased SWV. Both contribute through aerosol-induced tropopause warming; however, extreme wildfires reveal an additional self-lofting pathway that transports water vapour into the stratosphere. Complementary analysis of satellite observations and climate model simulations reveals an SWV enhancement of about 0.1 ppmv at 83 hPa, accumulating 76-203 million tons of water vapour during 2005-2021. This contribution explains 36 ± 7% of the observed SWV trend over this period, comparable to that from the global surface temperature increase. SWV changes induced by the surface temperature trend, moderate volcanic eruptions and extreme wildfire events have together effectively offset the sudden 10% SWV decrease observed around 2000. Episodic aerosol perturbations from moderate volcanic eruptions and extreme wildfires therefore emerge as a previously overlooked driver of SWV variability. Future projections of stratospheric composition, radiative forcing and ozone recovery should account for these aerosol-mediated processes, especially as extreme fires intensify in a warming world.
Glioblastoma is a lethal brain tumour for which current multimodal treatment rarely prevents recurrence. Therapeutic failure is driven by extensive intratumoural cellular heterogeneity with a microenvironment dominated b...Glioblastoma is a lethal brain tumour for which current multimodal treatment rarely prevents recurrence. Therapeutic failure is driven by extensive intratumoural cellular heterogeneity with a microenvironment dominated by tumour-associated macrophages that sustain tumour growth and immunosuppression. Although chimeric antigen receptor (CAR)-T cell therapies are being developed for glioblastoma, sustained response has been undermined by non-uniform antigen expression, antigen loss and microenvironmental barriers that are not directly engaged by tumour-targeting designs. These limitations motivate new strategies that address the disease as a coupled tumour-immune system rather than a single malignant compartment. Here we use a multi-omic target discovery platform to identify GPNMB as a dual-compartment antigen in glioblastoma. Anti-GPNMB CAR-T cells showed potent anti-tumour activity, with long-term disease control in orthotopic patient-derived xenografts and syngeneic glioma models through concomitant depletion of GPNMB tumour and immunosuppressive myeloid populations. By collapsing tumour control and microenvironmental reprogramming, these findings provide a new strategy for antigen selection and targeting in heterogenous, myeloid-rich solid cancers.
Fan S, Gao P, Huang K
… +22 more, Ying W, Xu L, Chen W, Ye H, Xi Y, Yang Y, Qian H, Li T, Tu B, Yuan H, Ma B, Wang Y, Zhong Z, Xiong J, Wang H, Kang L, Tang S, Chen X, Sun L, Xiang C, Li S, Qin P
Cells must rapidly counteract heat stress-induced hyperfluidization of the plasma membrane to prevent membrane damage, yet how cells achieve such early protection remains unknown. Here we show that in rice (Oryza sativa)...Cells must rapidly counteract heat stress-induced hyperfluidization of the plasma membrane to prevent membrane damage, yet how cells achieve such early protection remains unknown. Here we show that in rice (Oryza sativa), the P4-ATPase OsALA5, together with its β-subunit OsALIS2, mediates a heat-responsive flipping of saturated phosphatidylcholines that rapidly stabilizes plasma membrane fluidity. Using leaflet-resolved lipidomics and complementary transport assays, we demonstrate that heat exposure induces a minute-timescale shift in OsALA5 transport activity that leads to selective enrichment of saturated phosphatidylcholines in the cytoplasmic plasma membrane leaflet. This OsALA5-mediated saturated phosphatidylcholine flipping prevents plasma membrane hyperfluidization upon heat stress, thus mitigating ion leakage and cell death. Our analyses of OsALA5 orthologues in Arabidopsis thaliana and yeast support functional conservation of a rapid heat-associated response within a subset of plasma membrane-localized, phosphatidylcholine-transporting P4-ATPases. We identified a rare haplotype of OsALA5 that confers both heat tolerance and yield stability in multi-year, multi-location field trials. Thus, beyond identifying this P4-ATPase-mediated flipping of saturated phosphatidylcholines in response to heat stress and providing genetic resources to advance breeding of heat-tolerant crops, our study reveals how cells counteract heat stress-driven plasma membrane hyperfluidization at an earlier stage than the previously known transcription-dependent lipid remodelling response.
Intracellular bacterial symbioses have arisen myriad times in eukaryotes, with dozens known from insects alone. Beginning with Buchnera, the obligate endosymbiont of aphids, genomes of endosymbionts have illuminated thei...Intracellular bacterial symbioses have arisen myriad times in eukaryotes, with dozens known from insects alone. Beginning with Buchnera, the obligate endosymbiont of aphids, genomes of endosymbionts have illuminated their evolutionary origins and metabolic contributions to hosts. However, the mechanisms by which non-culturable endosymbionts enter host cells and suppress cellular immune processes have remained unclear. Here we show that an uncharacterized Buchnera protein, designated SyeA, was present in the Buchnera ancestor, is secreted into the host cytoplasm, is homologous to secreted effectors of bacterial pathogens and is essential for Buchnera transmission. Buchnera is transmitted through expulsion from specialized maternal cells and uptake by embryos. Using immunofluorescence microscopy, we found elevated SyeA levels after colonization of the embryonic cell, accompanied by actin accumulation at the entry site. SyeA localizes outside the host-derived membrane and actin layer surrounding each Buchnera cell within host cytoplasm. Knockdown of syeA expression disrupts colonization of embryos and embryonic development and elevates lysosomal activity, leading to Buchnera destruction. Our findings provide insights into how an anciently associated, mutualistic endosymbiont achieves its intracellular existence. SyeA represents a vestige of pathogenic origins that was followed by evolution of increased host control and erosion of the original, more complex pathogenicity machinery.
Yang ZL, Huang BC, Lin YK
… +16 more, Lin YR, Hsu HC, Chen HY, Wan Y, Tseng KW, Yang ST, Lo HC, Huang YJ, Zheng F, Yang N, Meng W, Min J, Huang PC, Lan YW, Li LJ, Chiu YP
Transistors based on two-dimensional (2D) materials are on the roadmap for the beyond 1 nm logic technology node. This stems from their ultrathin thickness and defect-free surfaces, granting remarkable electrostatic gate...Transistors based on two-dimensional (2D) materials are on the roadmap for the beyond 1 nm logic technology node. This stems from their ultrathin thickness and defect-free surfaces, granting remarkable electrostatic gate control. The physical channel length of 2D transistors may eventually reach <10 nm for advanced node devices. However, the equally important scaling limit for metal contacts remains unknown because of the lack of technology to directly probe the carrier injection region in contact areas. Here we use cross-sectional scanning tunnelling microscopy to directly measure the carrier transfer length as approximately 2.0 nm at the contact region of a bismuth-contacted monolayer MoS transistor. This approach allows contact scaling constraints to be determined, providing information for the development of future ultra-scaled electronic devices.
Chelicerates are a diverse group of terrestrial and aquatic arthropods, yet their origin and early evolution remain debated. Among different hypotheses, one proposes that the front appendages, known as chelicerae, of che...Chelicerates are a diverse group of terrestrial and aquatic arthropods, yet their origin and early evolution remain debated. Among different hypotheses, one proposes that the front appendages, known as chelicerae, of chelicerates evolved from the short great appendages (SGAs) of Cambrian megacheirans and that book gills originated from their trunk limbs. Although taxa such as Mollisonia plenovenatrix and Megachelicerax cousteaui provide important clues to the early evolution of chelicerates, the morphological transition from megacheiran-like appendages to true chelicerae and book gills remains unresolved. Here we use X-ray microtomography to reveal the three-dimensional anatomy of Urokodia aequalis, an early Cambrian euarthropod from the Chengjiang biota of China. Urokodia has a seven-segmented head with a sclerotized hypostome, pincer-like SGAs and biramous trunk appendages with overlapping exite flaps. Its pincer-like SGAs represent a bridge structure between the appearance of multisegmented SGAs and true chelicerae, and its trunk appendages support a megacheiran origin of book gills. Phylogenetic analyses consistently confirm the monophyly of Cheliceromorpha, with Urokodia being the earliest-branching upper stem-group chelicerate that links lower stem-group megacheirans to crownward forms such as Mollisonia and Megachelicerax.
Biodiversity change has elicited widespread concern over the consequences for functions and services provided by ecosystems. Despite extensive evidence for a positive effect of biodiversity on ecosystem functioning withi...Biodiversity change has elicited widespread concern over the consequences for functions and services provided by ecosystems. Despite extensive evidence for a positive effect of biodiversity on ecosystem functioning within a single trophic level, how this biodiversity effect varies with multi-trophic food web structure remains unresolved even though most ecosystems contain two to six trophic levels. We investigate how food web complexity modulates biodiversity-ecosystem functioning relationships in nature by quantifying energy fluxes as proxies for two principal ecosystem functions-primary consumption and predation-in 318 highly resolved, complex food webs from marine, lake, stream and soil ecosystems. Ecosystem functioning increased consistently with taxon richness across all trophic levels and ecosystems, which arose from greater vertical diversity (that is, maximum trophic level) and trophic complementarity of predators in more taxonomically diverse food webs. Furthermore, predator trophic complementarity increased predation fluxes in all freshwater ecosystem types. These findings highlight the threat of trophic downgrading to critical ecosystem functions (for example, biological control and maintenance of biodiversity and ecosystem stability) provided by predators, which are typically most vulnerable to anthropogenic disturbances. Our study demonstrates that the consequences of biodiversity change are deeply entangled within the web of life, emphasizing the need to conserve the trophic complexity underlying biodiversity-ecosystem function relationships.
Li JJ, Vasciaveo A, Karagiannis D
… +26 more, Sun Z, Gretarsson KH, Chen X, Ouerfelli O, Socciarelli F, Frankenstein Z, Dong H, Zou M, Yuan W, Yang G, Aizenman GM, Pannellini T, Xu X, Beltran H, Chen Y, Gardner K, Robinson BD, de Bono J, Gozani O, Abate-Shen C, Rubin MA, Loda M, Sawyers CL, Califano A, Lu C, Shen MM
Efficient priming of B cell precursors is a rate-limiting step in the induction of V2 apex broadly neutralizing antibodies (bNAbs). Here, we describe a novel germline-targeted HIV-1 Env (CAP256.OPT4) that increases the e...Efficient priming of B cell precursors is a rate-limiting step in the induction of V2 apex broadly neutralizing antibodies (bNAbs). Here, we describe a novel germline-targeted HIV-1 Env (CAP256.OPT4) that increases the efficiency of V2 apex bNAb precursor priming by 30-400 fold compared with wild-type HIV-1 Envs and induces - in >90% of macaques - neutralization breadth that includes N130-containing viruses. Using three different delivery platforms - persistently replicating simian human immunodeficiency viruses (SHIVs), protein nanoparticles, and mRNA - we show bNAb priming as early as 4 weeks post-infection or immunization, and neutralization breadth in plasma by 12 weeks. In 14 SHIV-infected macaques, neutralization breadth reached as high as 90% on a 21-virus panel with potency as great as 1:20,000 (50% inhibitory dilution, ID). Monoclonal bNAbs isolated from these animals were similarly broad and potent, with cryo-EM structures representing three distinct lineages revealing canonical needle-like HCDR3 binding. Env-Ab coevolution and structural analyses identified five key residues and loop features under positive selection and temporally associated with neutralization breadth. Importantly, prime-boost immunogens designed to capture these features induced broad and potent neutralization of globally diverse viruses including those containing N130 glycan. Further, rhesus bNAbs were not restricted to IGHD3-15*01 heavy chain alleles. These results expand the utility of the rhesus model for HIV-1 vaccine design and provide a molecular blueprint for inducing V2 apex bNAbs in rhesus and humans.
The high antigenic diversity of HIV has been a major obstacle to development of a broadly protective vaccine. Nevertheless, protective HIV broadly neutralizing antibodies (bnAbs) exist and have been proposed as templates...The high antigenic diversity of HIV has been a major obstacle to development of a broadly protective vaccine. Nevertheless, protective HIV broadly neutralizing antibodies (bnAbs) exist and have been proposed as templates for vaccine development. Germline-targeting is a conceptually radical vaccine design approach to elicit bnAbs, aiming to prime rare bnAb-precursor B cells possessing pre-determined human genetic and structural features shared with template bnAbs, and then guide B cell affinity maturation to potent bnAb evolution with heterologous boosters. Although the approach has shown promise in clinical and pre-clinical studies, it faces many immunological challenges and, to date, has not succeeded in generating bnAbs in humans or nontransgenic animals. Here, we report an adjuvanted protein germline-targeting vaccine tested in outbred nonhuman primates that generated bnAb-class memory B cells and sera capable of neutralizing diverse HIV clinical isolates. bnAb lineages were generated in ≥50% of animals, achieving up to 67% neutralization breadth compared to the reference bnAb. Vaccine-induced bnAbs exhibited precise structural mimicry of human bnAb interactions with HIV envelope (Env), matching the germline-targeting predictions. Furthermore, serum bnAb activity developed in 44% of animals and in the most striking instance reached titers expected to confer protection against diverse HIV isolates. These results demonstrate proof of principle that germline-targeting vaccines can reproducibly elicit prespecified classes of bnAbs to prespecified epitopes under endogenous conditions, supporting further optimization of this approach for HIV vaccine development.