Biology subjects

Zhang, Q.

Publications and source records attributed to Zhang, Q..

At least 109 records · Page 6Linked to original sources

bioRxiv2026

Mapping Individualized Dual-Axis Network Topology in Focal Epilepsy: Divergent Alterations in System Integrity, Integration, and Clinical Correlates

Focal epilepsy is increasingly recognized as a disorder of distributed brain systems, yet characterizations of patient-specific alterations in network organization that are clinically meaningful remain limited. Here, we used resting-state functional MRI to introduce an overlap-permitting individualized network-estimation framework anchored to normative references to derive two complementary, system-level axes of topology: (1) network correspondence, indexing the expression of canonical networks and the fidelity of intrinsic system boundaries, and (2) k-hubness, indexing cross-system integration and multi-functionality through multi-network participation. We used a large presurgical focal epilepsy cohort (305 patients, including temporal and extratemporal epilepsy; 224 healthy participants) and an independent multi-syndrome validation cohort spanning focal and common generalized and childhood epilepsy syndromes (903 patients; 666 healthy participants), to show: (1) correspondence disruption of both canonical and non-normative networks constitutes a broadly shared system-level endpoint that displays heterogeneity across individuals and differential impacts on cognition, (2) k-hubness reconfigurations reflect lateralized and syndrome-dependent redistributions of cross-system integration, demonstrating epilepsys impact on the functional multiplicity inherent to specific brain areas. These complementary axes shared variance yet showed dissociable clinical associations, with correspondence preferentially relating to neurocognitive deficits and k-hubness aligning with epilepsy clinical features. Accordingly, these axes demonstrate the presence of distinct topological phenotypes, providing clinically relevant, reproducible patient-level signatures that explain both the syndrome-specific and common effects of seizures on the organization and integration of canonical and individualized brain systems.

neuroscience↗

bioRxiv2026

InSTaPath: Integrating Spatial Transcriptomics and histoPathology Images via Multimodal Topic Learning

Spatial transcriptomic (ST) technologies enable the measurement of gene expression directly within tissue sections while preserving spatial context. Many ST platforms additionally generate paired histological images alongside spatially resolved transcriptomic profiles. However, most existing computational approaches only incorporate histology images as auxiliary features in representation learning models and typically produce latent embeddings that are difficult to interpret. We present InSTaPath (Integrating Spatial Transcriptomics and histoPathology images), a multimodal topic modeling framework that links transcriptional programs with tissue morphology. InSTaPath converts token-level embeddings extracted from pretrained histology foundation models into discrete image words through vector quantization, enabling histological morphology to be represented in a count-based form analogous to gene expression. InSTaPath then jointly analyzes image-word and gene expression counts to infer shared latent topics that are interpretable through both topic-gene and topic-image-word associations. Across multiple ST datasets, InSTaPath improves spatial domain identification and uncovers biologically meaningful relationships between gene programs and tissue morphology through pathway enrichment and in silico perturbation analyses.

bioinformatics↗

bioRxiv2026

APEX1 Protects Cardiomyocytes Against Oxidative Stress by Regulating p53 Protein Stability

Withdrawal statementThe authors have withdrawn this manuscript because the preprint was uploaded to bioRxiv before all co-authors had been fully informed and had provided their explicit consent to make the manuscript publicly available as a preprint. Following internal discussions, the authors have reached a consensus that they do not wish this version to be publicly available. We therefore kindly request that this preprint be withdrawn on the bioRxiv platform. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.

molecular biology↗

bioRxiv2026

Exploration of Orally Disintegrating Tablet for Sublingual Vaccination against Mucosal Bacterial Infection

Oral mucosal bacterial infections impose a substantial global disease burden, yet current clinical management typically reduces microbial load only transiently and rarely establishes durable protection at the oral surface. Analysis of 200 patients with periodontitis revealed that elevated levels of pathogen-specific salivary secretory Immunoglobulin A (sIgA) were strongly associated with reduced bacterial burden and improved clinical periodontal outcomes, identifying sIgA as a key determinant of effective oral protection. Guided by this observation, we developed a sublingual, orally disintegrating tablet vaccine (Capot) that incorporates bacterial extracellular vesicles providing a comprehensive repertoire of native antigens and multiple pathogen-associated molecular patterns, encapsulated within a calcium phosphate nanoshell to enable safe transmucosal delivery to submandibular lymph nodes. The rapidly disintegrating tablet format minimizes inadvertent swallowing and enhances local mucosal bioavailability. In mice and non-human primates, Capot induced robust and long-lasting salivary sIgA responses without overt oral mucosal or gastrointestinal inflammation and conferred strong protection against primary, recurrent, and antibiotic-resistant periodontitis. Together, these findings establish sublingual tablet vaccination as a practical strategy for selectively engaging oral mucosal immunity and preventing chronic bacterial diseases at oral mucosal surfaces.

bioengineering↗

bioRxiv2026

High-Fidelity Long-term Whole-embryo Lineage and Fate Reconstruction by Iterative Tracking with Error Correction

Reconstructing the complete cell lineages and fate maps of a living embryo has remained a central challenge in developmental biology. Here we introduce ITEC (Iterative Tracking with Error Correction), a fully unsupervised method that automatically reconstructs the lineage of every cell in the embryo with high fidelity. ITEC was validated with manually annotated lineages on four cross-species datasets including zebrafish, mouse, and Drosophila. We reconstructed the developmental lineages of a zebrafish embryo from terabyte-scale data of total 18.5 million cells with an estimated accuracy of over 99.7%. With ITEC, we revealed spatiotemporal dynamics of key morphogenetic processes such as somite boundary formation, demonstrated the various patterns of spatial sorting among adjacent organs or anatomical regions, and identified associations between cellular movement and spatial transcriptomics. ITEC provides a powerful platform for retrospective fate mapping and systematic exploration of developmental dynamics at the cellular scale and single-embryo level.

bioinformatics↗

bioRxiv2026

CHPT1-LCAT rewires lipolysis towards ferroptosis

Ferroptosis is driven by iron-dependent lipid peroxidation, yet how metabolic flux through central lipid pathways is selectively routed towards pro-ferroptotic lipid species remains unclear. Here, through an unbiased chemical-genetic screen targeting core metabolic enzymes, we identify diacylglycerol (DAG) as a licensing lipid intermediate whose pro-ferroptotic activity depends on its intracellular routing. Systematic manipulation of lipolytic flux reveals that ferroptotic vulnerability is not determined by bulk lipolytic output or downstream intermediates, but by the selective channelling of DAG into a distinct metabolic fate. Mechanistically, DAG is selectively routed through a previously unrecognized intracellular metabolic axis composed of choline phosphotransferase 1 (CHPT1) and lecithin-cholesterol acyltransferase (LCAT). We uncover an enzymatically active intracellular pool of LCAT (iLCAT) that cooperates with CHPT1 on Golgi-trans-Golgi network membranes to generate polyunsaturated cholesteryl esters that execute ferroptosis. Functionally, enforced DAG routing through this axis suppresses tumour growth via ferroptosis in vivo, whereas hepatocyte-specific inhibition of the CHPT1-iLCAT axis attenuates lipid peroxidation and disease progression in metabolic dysfunction-associated steatohepatitis. Together, these findings establish subcellular lipid routing, rather than lipid abundance per se, as a fundamental determinant of ferroptotic vulnerability.

cell biology↗

bioRxiv2026

Synergistic Inhibition of Notch Signaling and Forced Cell Cycle Re-entry Drive Müller Glia Reprogramming in Uninjured Mouse Retina

In regenerative species, such as teleost fish, Muller glia (MG) autonomously re-enter the cell cycle after injury and give rise to functional retinal neurons. In contrast, the loss of retinal neurons in mammals is irreversible due to the limited proliferative and regenerative ability of MG. Various strategies have been developed to induce proliferation of mature mouse MG with or without injury, yet most MG daughter cells retain glial cell fate. Here, we found that MG progenies maintain high Notch signaling, which may constrain their neurogenic potential. Conditional deletion of Rbpj, the central transcriptional effector of Notch, induced limited MG-to-neuron conversion in mature MG without proliferation. However, Rbpj deletion, combined with forced MG proliferation by overexpressing cyclin D1 and suppressing p27Kip1, significantly promoted MG dedifferentiation and ectopic expression of the neuronal marker Otx2 in MG daughter cells in uninjured mouse retina. Combining Notch inhibition with MG cell cycle re-activation not only increased the numbers of bipolar- and amacrine-like cells generated from MG but also promoted the further differentiation toward ON-cone, OFF-cone, and rod-bipolar subtypes. Single-nucleus RNA and ATAC sequencing data revealed that Notch inhibition facilitated the formation of MG-derived progenitor-like cells while MG proliferation increased chromatin accessibility of neurogenic genes. Notably, most MG-derived cells survived long term despite incomplete maturation. Together, our findings delineate how Notch inhibition and MG proliferation, alone or in combination, influence the regenerative potential of MG in the mammalian retina.

cell biology↗

bioRxiv2026

Volume regulation of cancer cells during osmotic pressure variation

Osmotic pressure is a fundamental physical determinant of cellular homeostasis, and its perturbation is recognized as a critical factor in cancer progression and therapy. However, whether cancer cells and normal cells respond to osmotic pressure changes differently remains unclear. Here, we subjected cancer and normal cells to osmotic shocks and compared their volume and traction force changes. Under hypotonic conditions, cancer cells recovered their volume and forces much more slowly than normal cells, although both cell types responded similarly to hypertonic shock. We found that actin-based cortical tension controlled the recovery speed; whereas myosin-mediated contractility tuned the volume change extent. Substrate stiffness also influenced the recovery process by altering cytoskeletal constraint. A theoretical model integrating adhesion energy, ion and water transport, and surface tension was developed. The results matched the experimental observations. Our work uncovered the distinct mechanobiological behaviors of cancer cells upon hypotonic shock, providing a mechanical framework for understanding cancer mechanisms that may inform the development of cancer therapeutics.

biophysics↗

bioRxiv2026

Hierarchical Gating of Cortical Population Dynamics Drives Pain

The prefrontal cortex and anterior cingulate cortex are key cortical hubs for pain regulation, yet the functional hierarchy between them remains unclear. We examined how the prelimbic cortex (PL) to ACC pathway regulates nociceptive processing and pain behavior in freely moving rats across synaptic, cellular, and network levels. Activation of the PL to ACC pathway reduced aversion to both evoked and spontaneous pain, whereas inhibition increased pain aversion, indicating that this circuit exerts tonic modulatory control. Meanwhile, ex vivo electrophysiology revealed that PL inputs recruit local ACC interneurons to produce feedforward inhibition of ACC pyramidal neurons. At the cellular level, in vivo microendoscopic calcium imaging showed that optogenetic activation of PL axon terminals suppresses nociceptive-evoked activity of ACC pyramidal neurons. At the network level, PL activation reduced pain-induced excitability while centralizing nociceptive information flow within the ACC, resulting in a gated, low output population state. Overall, these findings identify a hierarchically organized cortical circuit that tonically controls pain related sensory and affective experience.

neuroscience↗

bioRxiv2026

Lung microvascular rarefaction impairs pulmonary gas exchange and exacerbates heart failure with preserved ejection fraction

BackgroundDyspnea and exercise intolerance are the primary clinical symptoms of heart failure. Heart failure patients experience frequent hypoxemic episodes, yet underlying mechanisms and relevance remain poorly understood. In a cohort of heart failure patients and multiple animal models, we identify pulmonary capillary rarefaction driven by excessive autophagy in endothelial cells as a novel mechanism of hypoxemia and cardiac disease progression. MethodsA cohort of heart failure with preserved ejection fraction (HFpEF) patients was analyzed for parameters of left ventricular (LV) dysfunction and pulmonary gas exchange. Morphological and cellular mechanisms of impaired pulmonary oxygenation were assessed in three animal models of heart failure, namely two HFpEF models, SU5416-treated ZSF1 obese rats and high fat diet/L-NAME treated mice, and in rats subjected to aortic banding. Lung microvascular rarefaction was quantified by micro-computed tomography, stereology, flow cytometry and dye efflux. Cellular mechanisms of capillary loss were analyzed by single-cell transcriptomics, electron microscopy and immunofluorescence, and in mice with endothelial-specific deletion of the autophagy gene Atg7 (Atg7EN-KO). ResultsIn 234 HFpEF patients, advancing NYHA class was associated with progressive worsening of arterial oxygen saturation at rest and during exercise and a reduced lung diffusing capacity. Impaired gas diffusion correlated with indices of LV diastolic dysfunction. Impaired oxygenation and reduced exercise capacity were similarly evident in animal models of left heart disease, which showed a distinct loss of pulmonary microvessels and capillaries. Lung microvascular endothelial cells in HFpEF showed characteristics of increased autophagic flux and apoptosis. Relative to their wild type HFpEF controls, Atg7EN-KO mice had less capillary loss, restored normoxemia, improved exercise tolerance, and mitigated LV diastolic dysfunction. Additional studies in HFpEF mice corroborated the functional relevance of impaired gas exchange for the progression of left heart disease by demonstrating that additional hypoxia aggravated, whereas moderate hyperoxia improved LV function. ConclusionOur findings identify pulmonary microvascular rarefaction as a novel pathomechanism in heart failure that i) contributes to dyspnea and exercise intolerance, ii) impairs pulmonary gas exchange and iii) accelerates LV disease progression. Strategies targeting this axis such as moderate oxygen therapy may mitigate cardiopulmonary morbidity in heart failure. Clinical Trial RegistrationRegistered in the DRKS (Deutsches Register fur klinische Studien) as trial# DRKS00032974 at https://drks.de/search/en/trial/DRKS00032974.

physiology↗

bioRxiv2026

c-Fos enhances influenza virus replication by stabilizing the M2 protein and promoting autophagosome accumulation

The influenza A virus (IAV) employs multiple strategies to hijack host cellular machinery for efficient replication. While autophagosome accumulation is known to promote IAV replication, and the viral matrix protein 2 (M2) ion channel protein plays essential roles in viral uncoating, assembly, and autophagy induction, the mechanisms regulating M2 stability remain incompletely understood. Furthermore, although c-Fos participates in the replication of various viruses, its function in IAV infection has not been characterized. Here, we identify c-Fos as a critical proviral host factor that enhances IAV replication through stabilizing M2 protein and promoting autophagosome accumulation. We demonstrate that IAV infection triggers M2-mediated cytosolic calcium elevation, which upregulates c-Fos expression. The induced c-Fos physically interacts with M2 and prevents its proteasome and lysosomal degradation, thereby increasing M2 accumulation. This stabilized M2 protein as a viral protein directly facilitates viral replication while simultaneously promoting autophagosome accumulation to further enhance the process. Our findings reveal a novel positive feedback loop in IAV infection, establishing c-Fos as a key regulator of IAV replication through M2 stabilization, and highlighting these interactions as potential therapeutic targets for antiviral intervention. IMPORTANCEInfluenza A virus (IAV) remains a significant global health threat, causing substantial morbidity and mortality. Understanding virus-host interactions is crucial for developing antiviral strategies. Here, we uncover a previously unrecognized positive feedback loop in which IAV exploits the host transcription factor c-Fos to enhance its own replication. We demonstrate that IAV infection upregulates c-Fos through M2-mediated calcium signaling, and the induced c-Fos in turn stabilizes the viral M2 protein. This stabilized M2 not only directly supports viral replication but also promotes autophagosome accumulation, further facilitating virus production. Our findings establish c-Fos as a critical proviral host factor and reveal a mechanism by which IAV hijacks host cellular machinery to create a favorable environment for efficient replication. The identification of this c-Fos-M2 axis not only advances our understanding of IAV-host interactions but also opens new avenues for therapeutic intervention targeting this vulnerability in the viral life cycle.

cell biology↗

bioRxiv2026

Connectome lateralization in autism across the first 14 years: heterogeneity related to developmental stage, hemisphere, and pathophysiology

Hemispheric lateralization is fundamental feature to neurodevelopment, yet its dynamic evolution in autism across childhood and underlying molecular mechanisms remain unknown. Thus, using a large-scale resting-state fMRI dataset (n = 1553, ages 1-14), we investigated developmental heterogeneity in connectome lateralization. At the group level, abnormalities in dynamic connectome lateralization strength (DCLS) progressed from focal patterns in early childhood ([≤] 6 years) to widespread distributions by late childhood ([≤] 14 years), independent of symptom severity. The late-stage patterns were validated using the ABIDE dataset (n=153). Late childhood also showed marked increases in inter-individual heterogeneity. These alterations were significantly associated with clinical phenotypes, transcriptomic profiles, and multiple neurotransmitter systems, with distinct patterns across childhood groups. Our findings reveal a developmentally dynamic trajectory of connectome lateralization in autism and highlight late childhood as a critical window for stage- and individual-specific interventions tailored to substantial individual heterogeneity.

neuroscience↗

bioRxiv2026

Single-cell Analysis of Paired FFPE Placentas Reveals Trophoblast Reprogramming and Immune Dysregulation in Chronic Villitis of Unknown Etiology

Villitis of unknown etiology (VUE) is a common placental inflammatory lesion associated with adverse pregnancy outcomes, yet its mechanisms remain unclear. Mechanistic studies are hindered by reliance on archived formalin-fixed paraffin-embedded (FFPE) tissues, which are incompatible with standard single-cell methods. Here, we develop an FFPE-compatible single-nucleus RNA sequencing strategy and apply it to paired placentas from sequential pregnancies. VUE placentas exhibit pronounced immune expansion and disrupted trophoblast differentiation. A distinct extravillous trophoblast population and a subset of syncytiotrophoblasts aberrantly express classical MHC class I molecules, promoting natural killer cell engagement and indicating loss of trophoblast immune privilege. Pro-inflammatory maternal macrophages infiltrate, while fetal Hofbauer cells are reduced and transcriptionally reprogrammed. Increased intracellular viral burden and JAK-STAT signaling correlate with aberrant MHC class I expression, suggesting stress-induced local alloimmune activation. These findings define a cellular basis for disrupted maternal-fetal immune tolerance in VUE and demonstrate the utility of FFPE-based single-cell transcriptomics in placental disease.

cell biology↗

bioRxiv2026

Chemical Probing of Diacylglycerol Dynamics at Lipid Droplets

Diacylglycerols (DAGs) are central intermediates in lipid metabolism and signaling, yet their trafficking and persistence within lipid droplets (LDs) remain incompletely understood due to the lack of chemically stable, DAG-mimetic imaging tools. Here, we report the development of a family of solvatochromic fluorescent lipid analogs, termed DONDI, designed to probe DAG-associated dynamics at LDs. These probes are based on a 1,8-naphthalimide scaffold conjugated to modified aminoglycerol backbones bearing oleoyl chains to mimic native glycerolipids. Biophysical characterization and atomistic molecular dynamics simulations revealed probe-specific membrane insertion and hydrogen-bonding behaviors consistent with distinct lipid-mimetic properties. Live-cell imaging in NIH 3T3 fibroblasts demonstrated that DONDI probes were efficiently internalized and selectively accumulated within lipid droplets. Structure-function analysis identified DONDI-5 as the closest mimic of 1,2-diacylglycerol, displaying rapid uptake, strong LD enrichment, and prolonged intracellular retention without detectable relocalization to other cellular membranes. These properties enabled sustained visualization of LD-associated DAG pools over extended time scales. Collectively, this work establishes DONDI-5 as a chemically stable DAG-mimetic probe and provides direct experimental support that DAGs can be transported to and transiently stored within lipid droplets without prior conversion to triacylglycerols.

biochemistry↗

bioRxiv2026

Sex-specific Genetic Regulatory Effects in Chickens

Sexual dimorphism is a defining vertebrate feature, yet its sex-specific molecular architecture remains poorly understood. Here we established a sex-balanced, uniformly reared chicken cohort to map this landscape, integrating individual whole-genome sequencing with 7,969 bulk and 779,380 single nucleus transcriptomes across 32 tissues from 280 birds. We identified 495,098 independent expression quantitative trait loci for 20,194 genes, including 10,937 loci modulated by cell-type composition. Notably, 340 genes were regulated by 449 loci in a sex-dependent manner, significantly enrichment in endocrine tissues like adipose and the adrenal gland. Furthermore, we fine-mapped 1,219 structural variants, demonstrating their unique roles to tissue- and sex-specific expression beyond SNPs. Ultimately, we showed the utility of these regulatory effects in elucidating the molecular basis of metabolism and complex traits in both chickens and humans. This comprehensive atlas of regulatory effects provides profound insights into the genomic and molecular basis of sexual dimorphism in vertebrates.

genomics↗

bioRxiv2026

Retrotransposons Promote Clonal Hematopoiesis Through Aging-Related Stromal Inflammation

Clonal hematopoiesis (CH) is an age-related phenomenon driven by the expansion of mutant hematopoietic stem cell (HSC) clones, most commonly harboring mutations in DNMT3A. While inflammation is known to promote CH, the upstream triggers of this inflammatory state remain unclear. We show that aging selectively upregulates retrotransposons in the non-hematopoietic cell compartment of the murine bone marrow, particularly in mesenchymal stromal cells. Using in vivo competitive transplant models, we demonstrate that retrotransposon-induced inflammation cell-extrinsically promotes Dnmt3a-mutant HSC expansion. This competitive advantage arises from mutant HSC resistance to inflammation-driven cell cycle perturbations. Mechanistically, we show that retrotransposon activation induces type I interferon signaling via viral mimicry, such that stromal knockdown of either Irf3 or Sting abrogates the competitive advantage of Dnmt3a-mutant HSCs. Our findings establish stromal-selective retrotransposon reactivation as a previously unrecognized, non-cell-autonomous source of inflammation that contributes to age-associated CH. Key PointsO_LIRetrotransposons remodel the aging niche through viral mimicry pathways to promote Dnmt3a-mutant clonal hematopoiesis. C_LIO_LIRetrotransposon-mediated inflammation impairs wild-type HSC fitness, while Dnmt3a-mutant HSCs resist inflammatory changes. C_LI

cancer biology↗

bioRxiv2026

Wildfire emitted particulate matter induces ovarian hyperandrogenism through aryl hydrocarbon receptor activation

Wildfires have become more frequent and intense worldwide. Wildfire emitted particulate matter (WFPM) can be more toxic than urban background PM due to its greater content of nanoscale size (WFPM0.1) and presence of more polar organic compounds, including polycyclic aromatic hydrocarbons (PAHs). While exposure to WFPM has been linked to cardiovascular and respiratory diseases, its impact on female reproduction remains elusive. Here, we used an in vivo mouse intratracheal exposure model and a 3D ovarian follicle culture system, together with molecular, transcriptomic, and computational approaches, to examine the female reproductive effects of lab-synthesized (LS-WFPM0.1) and real-world Canadian WFPM0.1 (C-WFPM0.1), collected from the New York City and New Jersey metropolitan area during the June 2023 wildfire events. Intratracheal exposure to environmentally relevant dose of LS-WFPM0.1 disrupted mouse estrous cycles and elevated serum concentrations of estradiol and testosterone. RT-qPCR and single-follicle RNA-sequencing (RNA-seq) analysis revealed altered steroidogenic genes, transcriptomic changes, and activation of aryl hydrocarbon receptor (AhR) in antral follicles from mice treated with LS-WFPM0.1. LS-WFPM0.1 consistently increased testosterone secretion and stimulated genes related to androgen synthesis and AhR in vitro. Single-follicle and single-oocyte RNA-seq analysis identified differentially expressed genes related to inflammation in somatic cells and mitochondrial respiratory chain in oocytes. Both C-WFPM0.1 and benzo[a]pyrene, a high-molecular-weight PAH, reproduced these ovarian defects. Mechanistically, AhR inhibition reversed hyperandrogenism induced by WFPM0.1. Together, our findings suggest that WFPM0.1, an increasingly pervasive environmental exposure, adversely impacts female reproductive functions by disrupting ovarian steroidogenesis and inducing hyperandrogenism through AhR activation, highlighting an urgent unmet need for further mechanistic studies and epidemiological investigations to define the reproductive risks of wildfire smoke exposure in human populations.

pharmacology and toxicology↗

bioRxiv2026

Unwinding of RNA G-quadruplexes induces mouse and human totipotency

Totipotent stem cells (TotiSCs) have significant application prospects in regenerative medicine and assisted reproductive technologies. The mRNAs of totipotency-related genes are rich in G-quadruplex (G4) structures, while the G4 density of pluripotency-related genes mRNAs is relatively low, suggesting a potential functional link between RNA G4s and the totipotent state. Nevertheless, how G4s are involved in the establishment or maintenance of mouse and human totipotency remains poorly understood. Here, we demonstrate that FC3, a small-molecule RNA G4 unwinder, enables efficient induction and maintenance of mouse and human TotiSCs (mG4TotiSCs and hG4TotiSCs). These FC3-induced TotiSCs closely recapitulate the molecular features of 2-cell-stage blastomeres via transcriptomic and epigenomic profiling. Functionally, both mG4TotiSCs and hG4TotiSCs exhibit totipotent capacity: they differentiate into embryonic and extraembryonic lineages in vitro and in chimeric embryos; moreover, they autonomously self-organize into blastocyst-like structures without exogenous signalling. Mechanistically, we identify that Gata2 mRNA, a G4-containing transcript, is essential for FC3-mediated reprogramming. Genetic disruption of Gata2 would abolish totipotency induction. Collectively, RNA G4 unwinding is a new regulatory axis governing totipotency acquisition and maintenance, providing insight into the functional interplay between nucleic acid secondary structures and cell fate plasticity.

cell biology↗