Simultaneously introducing diverse genomic edits remains a challenge in crop genome engineering. Here we describe a twin prime editing-based knockout (TKO) system that installs stop codon clusters (SCCs) for precise translational termination with minimal in-frame mutations. TKO achieves knockout efficiencies of up to 70.5%, 58.6% and 75.1% in rice, maize and wheat protoplasts, respectively, and produces heritable knockout alleles in 96.8% of regenerated rice plants. In hexaploid wheat, TKO outperforms Cas9 4.2-fold in generating triple-homolog knockouts, largely by reducing in-frame mutations. Orthogonal TKO editors with sequence-divergent SCCs enable simultaneous knockout of up to ten genes without cross-interference. Integration of TKO with conventional prime editing establishes TRIM1 (TKO editor-enabled gene rupture and development of integrated multitype genome modification system) for simultaneous knockout and precise editing, achieving a 22.8% coediting of four genes in rice. TRIM2 extends this capacity to kilobase-scale modifications through a prime editor-recombinase system, enabling a 4.9-kb insertion (1.2% efficiency) and gene knockout (up to 79.8%) in protoplasts.
Achieving efficient and precise replacement of large genomic fragments with isogenic sequences remains a major challenge in plant genome editing, limiting the exploitation of natural allelic diversity for trait improvement. This study aims to develop an optimized prime editing (PE) strategy for high-efficiency, large-fragment isogenic sequence replacement (ISR) in rice. We systematically compared nuclease-based PE, template-jumping PE (TJ-PE), and GRAND PE strategies. We engineered a series of M-MLV reverse transcriptase (RT) variants and evaluated their performance in rice protoplasts and stable transgenic lines. TJ-PE outperformed other strategies in ISR efficiency and precision. Engineering the Moloney murine leukemia virus reverse transcriptase yielded rPE14e4 (T128N/D200C/V223Y/L435K), which enhanced ISR efficiency by 4.5-fold and enabled precise replacements up to 250 bp. We also discovered that unintended microhomology between primer binding site (PBS) and reverse transcription template (RTT) can cause on-target byproducts, and its disruption improves editing fidelity. Applying the optimized rPE14e4-TJ-PE system, we successfully rewrote a 174-bp coding region of the xa10 gene in the elite rice cultivar N9208. We established an efficient PE-mediated system for large-fragment ISR in rice. The optimized strategy and engineered RT variant significantly expand the capability for precise gene rewriting, accelerating functional genomics and molecular breeding in crops.
Multiplex editing is crucial for analyzing complex multiple-gene traits in woody plants, yet its application remains limited by low transformation efficiency and lengthy regeneration cycles. To overcome these barriers, this study establishes an efficient protoplast isolation protocol for Pyrus, employing 1.0% cellulase R10 and 0.4% macerozyme R10 with an 8.5 h digestion, and demonstrates its broad applicability across seven economically important woody plants. Coupling a 40% PEG-4000-mediated transfection regimen with DNA-free CRISPR/Cas9 ribonucleoprotein (RNP) delivery enabled multiplex genome editing in isolated protoplasts. Using this platform, simultaneous disruption of PbrARC3, PbrPARC6, and PbrFtsZ2-1a, key components of the chloroplast division apparatus, consistently reproduced macro-chloroplast abnormalities, confirming effective multigene perturbation within a single cellular context. Notably, chloroplast division failure activated chloroplast-to-nucleus retrograde signaling, evidenced by induction of nuclear stress-response genes PbrRBOHD and PbrZAT12, a concomitant surge in reactive oxygen species, and progression to severe cellular deformation. These results establish a rapid, cross-genus protoplast-RNP workflow that enables DNA-free multiplex editing and accelerates genotype-to-phenotype analyses in woody perennials. The approach provides a practical foundation for functional genomics and supports advances in non-transgenic precision breeding of tree crops.
Bone regeneration remains constrained by incomplete osteogenic commitment of mesenchymal stem cells (MSCs), underscoring the need for precise lineage control. CRISPR/Cas-based epigenome editing provides programmable access to chromatin regulators without altering the DNA sequence, and catalytically inactive Cas9 (dCas9) fused to transcriptional activators, repressors, or chromatin modifiers enables locus-specific modulation of key osteogenic networks, including RUNX2, OSX, and BMP2, while suppressing inhibitory loci such as PPARG, SOST, and DKK1. Multiplex strategies further allow the concurrent activation of osteogenic genes and repression of adipogenic or Wnt antagonists, reshaping lineage allocation in vitro and in vivo. Delivery innovations-from AAV vectors and lipid nanoparticles to biomaterial scaffolds and extracellular vesicles-support local and systemic applications with increasing precision, while whole-genome chromatin profiling and high-fidelity Cas variants reduce off-target risk, and CRISPRoff/on platforms provide reversible and heritable control of transcriptional states. Proof-of-concept studies in small animals demonstrate bone repair in preclinical models, with emerging large-animal data highlighting translational potential. Remaining challenges include payload size, immunogenicity, durability of epigenetic states, GMP-grade manufacturing, and regulatory classification. Looking ahead, advances such as AI-guided gRNA libraries, mechano-responsive scaffolds, and long-term tracking of epigenetic memory may yield durable "smart" osteo-epigenetic therapies. Collectively, CRISPR/dCas9-based epigenome editing is progressing from mechanistic exploration toward clinically viable strategies for skeletal regeneration.
Becker Muscular Dystrophy (BMD) is caused by in-frame mutations in the DMD gene, leading to the production of internally truncated but partially functional dystrophin. Although cardiac involvement is a major contributor to disease burden in BMD, the cellular mechanisms driving cardiomyopathy remain incompletely understood. While emerging evidence suggests that iron imbalance may contribute to oxidative stress and mitochondrial dysfunction in muscular dystrophies, its role in BMD-associated cardiomyopathy has not been defined. Building on our previous findings of dysregulated iron homeostasis in dystrophin-deficient cardiomyocytes from Duchenne muscular dystrophy (DMD), we investigated whether similar alterations are present in BMD using patient-specific and genome-corrected hiPSC-CM models. HiPSC lines derived from two BMD patients and their CRISPR/Cas9-corrected isogenic controls displayed normal karyotype, pluripotency, and efficient differentiation into cardiomyocytes (hiPSC-CMs). BMD hiPSC-CMs showed elevated ROS levels and decreased cytoplasmic and mitochondrial labile iron pools, accompanied by reduced expression of mitoNEET (CISD1), a regulator of mitochondrial iron handling. We also detected changes in the expression of genes involved in iron storage (FTH1), uptake (TFRC), and export (SLC40A1), suggesting a dysregulation of iron trafficking. Importantly, correction of DMD mutation by CRISPR/Cas9 gene editing reversed the effects observed in BMD cardiomyocytes. These results extend our previous observations in DMD to BMD cardiomyocytes and suggest that full-length dystrophin is essential for maintaining cardiac iron homeostasis.
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Prime editing (PE) enables precise nucleotide changes but has limited utility in high-throughput functional screening due to low editing efficiency. Here, we developed EvoPRIME, a PE3-based screening platform that integrates Csy4-mediated processing of Pol II-driven guide RNAs with a fluorescence-based enrichment strategy. EvoPRIME achieves comparable efficiency to state-of-the-art prime editing systems without requiring mismatch repair (MMR) suppression. We validated the performance of EvoPRIME through a loss-of-function (LOF) dropout screen targeting essential genes, which showed improved sensitivity and reproducibility, and a gain-of-function (GOF) saturation mutagenesis screen of the EGFR tyrosine kinase domain under osimertinib selection, which identified both known and uncharacterized resistance-conferring mutations. Leveraging EvoPRIME, we further conducted a high-throughput screen that functionally assesses both gain- and loss-of-phosphorylation (GOP and LOP) mutations. Among the hits, GAB1 S419E emerged as a phosphomimetic mutation that activates AKT signaling and confers osimertinib resistance. These studies establish EvoPRIME as a versatile platform for uncovering functional variants.
Drought at the reproductive phase, referred to as terminal drought is a major constraint which severely limits seed yield in chickpea. Development of drought tolerant cultivars entails identification of key genes which govern drought tolerance. In order to identify such genes, two different genotypes, which contrast for drought stress tolerance were analysed under terminal drought. The drought tolerant (DT) variety, ICC 4958 and the drought sensitive (DS) variety, ICC 1882 responded differently to terminal drought stress. To identify the drought-induced changes in gene expression, the root transcriptomes of both genotypes were analysed. Two genes belonging to the SPL (SQUAMOSA promoter binding protein-like) transcription factor (TF) family, CarSPL1 and CarSPL9 were significantly (p < 0.05) upregulated, by a fold change of more than 8, in the sensitive genotype relative to the tolerant genotype. Many stress responsive genes like HKT1;3-like, NAC57, GolS3, WSD11, LTP4, etc. were downregulated in the sensitive genotype compared to the tolerant genotype. The CarSPL1 and CarSPL9 proteins shared a high degree of homology with the CaSBP13 and OsSPL6 of pepper and rice, respectively, which are known negative regulators of drought stress response in these crops. In silico analysis revealed a high amenability of the two genes for gene editing as high-efficiency sgRNAs (single guide RNAs) with high (> 66) out-of-frame scores, could be designed. The findings suggest that the identified CarSPLs act as negative regulators of drought tolerance in chickpea, allowing their utilization as potential targets for gene editing to engineer drought stress tolerance.
Alternative splicing (AS) is a fundamental RNA processing mechanism, which generates different RNA transcripts and consequently different protein isoforms from a single gene. This increases the diversity of proteins within an organism and can fine-tune biological processes. This review examines how cardiac-enriched RNA-binding proteins establish heart-specific splicing programs governing aspects of cardiac development, function, and disease. Developmentally, coordinated sarcomeric isoform switches underpin the foetal-to-adult transition and further isoform rewiring in ion channel and kinase genes determine electrophysiology and excitation-contraction coupling. AS contributes to the pathogenesis of several cardiomyopathies and emerging datasets suggest that pathological hypertrophy engages distinct splicing signatures compared with physiological hypertrophy. This review summarizes diagnostic and prognostic opportunities arising from bulk, long-read, and single-cell/nucleus transcriptomics, which resolve cell type-specific isoforms and disease-associated switches. Circulating RNA biomarkers (including splice ratios and circularRNAs) may signify myocardial remodelling and arrhythmic risk. Integrative approaches that link AS with proteomics and genomics improve variant interpretation, reveal previously unannotated protein isoforms, and enable tracking of disease progression and therapy response. Finally, an outline of therapeutic strategies to modulate AS in cardiovascular disease (CVD), including antisense oligonucleotides, small molecules, and genome-editing modalities (CRISPR, base, and prime editing), is provided. The major challenges that remain before splice-targeting therapeutics can be targeted to treat cardiovascular disease are highlighted. Lessons from neuromuscular indications establish clinical feasibility of splicing correction and motivate translation to cardiology. Together, mechanistic insight, biomarker development, and therapeutic innovation position RNA splicing as a tractable axis for precision cardiovascular medicine.
To evaluate the laboratory cleaning efficacy of six commonly used non-invasive methods for removing stains from occlusal fissures before caries diagnosis. 60 extracted, caries-free permanent molars with visible occlusal staining were divided into six groups (n= 10): AquaCare air abrasion, Kerr prophylactic paste, Whiteness HP (35% H₂O₂), HealOzone, oxygenated water + pumice, and Opalustre. All samples were analyzed pre- and post-treatment using a dental microscope, Diagnodent Pen, QLF imaging (Qraypen), 3D Geomagic Control X software for volumetric evaluation and photo editing software for pixel evaluation. Statistical analyses were performed with Kruskal-Wallis and the Friedman's two-way ANOVA test (P< 0.05). All methods significantly reduced surface discoloration (P< 0.05). AquaCare showed the least residual staining, while HealOzone had the highest (P= 0.043). Volumetric analysis revealed no intergroup enamel loss (P> 0.05), although a significant change was noted in the Whiteness HP group (P= 0.017). QLF parameters (ΔF and ΔQ) improved significantly in Aquacare, HealOzone, and Opalustre (P< 0.05). Diagnodent Pen readings decreased significantly in the Aquacare, Kerr paste, HealOzone, and oxygenated water + pumice groups (P< 0.05). All tested methods effectively removed fissure stains without causing measurable enamel loss. HealOzone, Aquacare, and Opalustre demonstrated the most balanced performance, combining high cleaning efficacy with minimal surface alteration. The findings emphasize the value of multimodal, minimally invasive approaches for optimizing diagnostic accuracy in occlusal fissure caries detection. When making a caries indication, the staining of the fissures may affect the accuracy of the indication, but the most minimally invasive method should be preferred to clean the stained fissures.
Oncogenic viruses cause approximately 20% of cancers globally burden, with Epstein-Barr virus and high-risk Human papillomavirus recognized as major contributors to epithelial malignancies. Increasing evidence suggests that EBV-HPV co-infection may enhance tumour progression through overlapping molecular and immunological mechanisms, particularly in cervical, oropharyngeal, and nasopharyngeal cancers. This review critically summarizes current evidence regarding the cooperative role of EBV and HPV in carcinogenesis while distinguishing viral co-presence from biologically active co-infection. EBV latent proteins, including LMP1, LMP2A, and EBNA1, activate oncogenic signalling pathways such as NF-κB, PI3K/Akt, and JAK/STAT, whereas HPV oncoproteins E6 and E7 disrupt p53 and retinoblastoma (Rb) tumour suppressor pathways. Together, these alterations may promote genomic instability, chronic inflammation, immune evasion, epigenetic dysregulation, and epithelial-mesenchymal transition (EMT), thereby enhancing invasive and metastatic potential. Epidemiological studies report higher frequencies of EBV-HPV co-detection in advanced lesions and aggressive tumours; however, causal synergy remains insufficiently validated because of methodological heterogeneity and variability in viral detection techniques, including PCR, in situ hybridization, and immunohistochemistry. Emerging technologies such as spatial transcriptomics and single-cell profiling may improve characterization of biologically meaningful co-infection. In addition, circulating viral DNA, viral microRNAs, and HPV genotyping are being explored as biomarkers for disease monitoring and prognosis. Therapeutic strategies targeting viral oncogenes, immune checkpoints, and gene-editing technologies also represent promising investigational approaches. Overall, EBV-HPV co-infection represents a biologically plausible but incompletely understood contributor to tumour aggressiveness, emphasizing the need for standardized diagnostics, longitudinal studies, and functional experimental models.
Poly-γ-glutamic acid (γ-PGA) is a natural biopolymer with broad application potential. Molecular weight (MW) is a key physicochemical parameter governing its structural properties, functional performance, and application scope. Recently, sustainable biosynthesis of γ-PGA with controllable MW has gained attention because of its environmental sustainability and process flexibility. With advances in molecular editing and synthetic regulation, MW-control strategies have shifted from random mutagenesis-based strain improvement to precise gene-engineering regulation. This review surveys natural microbial resources producing γ-PGA with diverse MWs and engineering strategies enabling de novo γ-PGA biosynthesis in multiple microbial chassis. To address limited production efficiency, we summarize metabolic engineering approaches for improving γ-PGA yield and MW tunability, including precursor supply optimization, carbon-flux redistribution, transcriptional regulation, use of non-food renewable substrates, and mitigation of metabolic burden from multi-layered engineering. To promote customized production of low-MW γ-PGA, we highlight hydrolase-centered strategies, emphasizing hydrolase screening, optimization of expression elements, and coordinated regulation of γ-PGA stereochemical configuration. Finally, we review applications of γ-PGA with different MWs in food, biopharmaceutical, and agricultural sectors, critically examine links between molecular characteristics and application requirements, and discuss future functional diversification and industrial-scale development. The increasing scientific and industrial interest in γ-PGA stems from its versatile functional properties across agricultural, food, and industrial applications, in which MW is a key determinant of both biological activity and physicochemical behavior. This review systematically examines metabolic engineering strategies that enable the de novo design and customized biosynthesis of γ-PGA through coordinated modulation of synthase–hydrolase regulatory networks, within an integrated framework encompassing synthesis, modification, and application. The mechanistic insights presented here provide a rational roadmap and practical guidance for researchers advancing microbial biopolymer engineering.
The CRISPR/Cas9 system enables precise and efficient genome editing, but its efficacy heavily relies on sgRNA activity. Although deep learning has been widely applied to sgRNA activity prediction, existing methods often integrate multiple biological features without a well-designed fusion strategy. To tackle this issue, we present CrisprFusion, a deep learning framework that explicitly encodes four biological features through a four-branch input structure. The core of our model is a novel Multi-Grain Cross Attention Fusion Module, which performs fusion at two levels: branch-level gating for adaptive reweighting of different modalities, and token-level alignment for capturing position-specific interactions along the 23-nt sgRNA sequence. We evaluate CrisprFusion on seven high-throughput datasets with six representative baselines. Our method achieves consistent and superior average performance across all datasets and remains competitive in cross-cell-line validation on four functional screens. Ablation experiments verify the effectiveness of the proposed fusion module, and attention visualization reveals the importance of individual biological features. Overall, CrisprFusion offers an effective and interpretable approach for multimodal biological feature integration in sgRNA activity prediction.
Polybromo 1 (PBRM1), encoding the BAF180 subunit of the polybromo-associated BAF (PBAF) chromatin-remodeling complex, is commonly lost or mutated across malignancies. Despite its prevalence, tailored therapeutics for PBRM1-defective cancers remain limited. PBRM1 loss is associated with elevated replication stress and DNA damage responses, implying dependence on compensatory repair pathways. Given the lack of genotype-matched drugs, exploiting DNA-repair dependencies may provide a precision option for PBRM1-deficient disease. We pursued a synthetic-lethality strategy in colorectal cancer to test whether clinically used PARP inhibitors selectively suppress PBRM1-deficient cells and to define the linked cell-cycle and stress-response mechanisms. We also compared PARP inhibition with broader chromatin-remodeler targeting. Isogenic PBRM1-/- HCT116 colorectal carcinoma cells were generated by CRISPR/Cas9 lentiviral editing using sgRNAs cloned into lenti-CRISPR-V2. Knockout was confirmed by Western blotting, Sanger sequencing, and RT-qPCR. A focused compound screen compared four agents PARP inhibitors olaparib and rucaparib, the multi-target chromatin remodeler inhibitor AU-24,118, and the SMARCA2/4-targeting degrader AU-1530 using dose-response CCK-8 viability assays and selectivity indices. We then validated our results by 12-day colony-formation assays. Mechanistic analyses measured drug-induced G2/M accumulation by propidium iodide staining and flow cytometry and quantified apoptosis by Annexin V/PI dual staining, with significance assessed by t-test or two-way ANOVA. PBRM1 loss confers selective hypersensitivity to PARP inhibitors, which intensify DNA-damage signaling, promote G2/M checkpoint arrest, trigger apoptosis, and induce stress-response genes such as CSRNP3. Although this effect appears context-dependent and was not observed uniformly across all PBRM1-/- models tested. These results support further evaluation of PBRM1 as a potential predictive biomarker in defined molecular contexts rather than as a universal marker of PARP inhibitor sensitivity.
P2X receptors (P2XRs) are ATP-gated cation channels that play a pivotal role in chronic visceral pain (CVP). This review highlights the central contribution of the ATP-P2X3/4/7 axis in peripheral and central sensitization underlying CVP. Recent discoveries have identified the non-coding RNA miR-1306-3p as an endogenous nanomolar agonist of P2X3 receptors, coupling chronic stress to visceral pain via an epigenetic pathway. Moreover, persistent DNA methylation changes at the P2RX7 locus in spinal astrocytes create a "pain memory" that limits the durability of conventional antagonists. The first-generation P2X3/P2X7 antagonists (e.g., AF-219, AZD-9056, NC-2600) failed in clinical trials, primarily due to species-specific receptor pharmacology, the lack of ATP-based biomarkers for patient stratification, and irreversible central sensitization driven by epigenetic marks. To overcome these hurdles, we propose a precision-medicine framework that includes: (1) CRISPR-dCas9-based epigenome editing as a potential one-time "pain-memory eraser"; (2) patient stratification using sweat-ATP levels; and (3) human iPSC-derived neuron screening to improve translational predictability. This integrated approach holds promise not only for CVP related to irritable bowel syndrome (IBS), but also for other pain conditions.
Multidrug resistance (MDR) in bacteria poses a significant global threat to public health. Elucidating the core molecular regulatory mechanisms underlying MDR is crucial for developing novel intervention strategies. In Gram-negative bacteria, the phage-derived Type VI Secretion System (T6SS) functions as a versatile "molecular weapon". Beyond its classical role in interbacterial antagonism, T6SS acts as a key indirect regulatory hub for modulating bacterial antimicrobial resistance (AMR) in a strain-specific and environment-dependent manner. Although T6SS does not directly participate in the expression of antibiotic resistance genes (ARGs) or the catalytic activity of AMR-related enzymes, it profoundly influences the development and dissemination of AMR across strains and species through multiple indirect mechanisms. This review systematically analyzes four core T6SS-mediated mechanisms: (1) secretion of AMR-associated effectors and biofilm modulation to establish resistant phenotypes; (2) formation of synergistic regulatory networks with biofilm development, oxidative stress response, efflux pumps, and other secretion systems, which specifically enhances bacterial antibiotic tolerance (distinct from antibiotic resistance phenotypes); (3) acceleration of horizontal gene transfer (HGT) of ARGs through natural transformation, plasmid conjugation, and outer membrane vesicle (OMV)-mediated transport; (4) targeted interbacterial killing enabling antimicrobial-resistant strains to overcome colonization resistance, gain ecological advantages, and exacerbate clinical infections. Building on this framework, novel anti-AMR strategies targeting T6SS are outlined, including direct disruption of T6SS assembly and function, interference with upstream regulators (e.g., quorum sensing), optimization of CRISPR-Cas gene editing, and engineered T6SS-targeted delivery platforms. By dissecting the T6SS-driven AMR network and its clinical translational potential, this review provides a foundation for designing next-generation therapies to reverse AMR and block ARG transmission and also discusses existing bottlenecks limiting the clinical translation of T6SS-targeted therapies, while identifying critical future research directions such as deciphering species-specific mechanisms and enhancing targeted delivery efficiency.
Pentatricopeptide repeat (PPR) proteins are central regulators of organellar RNA metabolism, mediating RNA editing, splicing, stabilization, and translation in chloroplasts and mitochondria. Because organellar gene expression requires fine-tuned coordination between nuclear and cytoplasmic genomes, hybridization may disrupt cytonuclear balance. However, whether essential nuclear-encoded organellar regulators such as PPR proteins undergo regulatory divergence in natural hybrid genomes remains unresolved. In this study, a comprehensive genome-wide and transcriptome-integrated analysis of PPR genes was performed in wild Prunus yedoensis. Using a haplotype-resolved reference genome, coverage-based parental-origin classification, multi-tissue RNA-seq profiling, structural subclass annotation, organelle-targeting prediction, and alternative splicing analysis were integrated. Among 992 curated PPR genes, coverage-based parental-origin composition was comparable to the genomic background, with no enrichment of maternal- or paternal-origin genes. This pattern was broadly conserved across P- and PLS-type subclasses, despite modest but statistically significant differences in parental-origin composition. Expression profiling revealed highly overlapping expression distributions and comparable tissue-specificity (τ) values among parental-origin categories. Moreover, no statistically significant tissue-dependent alternative splicing (AS) events were detected for PPR genes. Although a limited subset exhibited tissue-associated expression enrichment, homology-based functional annotation indicated conserved roles in organellar RNA processing. Collectively, these results indicate that PPR genes constitute a transcriptionally constrained regulatory module that preserves organellar functional stability in hybrid genomes, underscoring the stability of essential cytonuclear regulatory systems.
Currently, in vitro models of microvascular biology rely on self-assembly of vascular cells in compatible gels. However, the stochastic nature of this process results in large variations in lumen sizes, perfusion continuity, and shear stresses, making systematic and reproducible analysis challenging. Here, we report a new technology to generate artificial capillaries on a chip with custom control over lumen sizes and architectures using a combination of femtosecond laser cavitation and collagen casting within multi-chambered microfluidic chips. The design allows seeding of endothelial cells within capillary-sized microchannels and seeding of stromal cells within top-open silos, with independent control over seeding sequence and media compositions. Results show that endothelialized microchannels, coined as artificial capillaries, exhibit excellent barrier function with reproducible control over lumen sizes (ϕ=8-40µm) and their architectures (straight, curvatures, tapered, branched). The physical flow parameters measured across the lumen (namely, flow shear) and at the channel outlets (flow velocities) have been validated against high-fidelity numerical assessments from the Large Eddy Simulation scheme within the digitized versions of microchannels. The experiment-computation compatibility enabled us to predict changes in regional velocity and wall shear stresses within artificial capillaries for various capillary architectures. We also show that in situ editing of artificial capillaries in the form of adding new branches or adding occlusions is possible. Lastly, we developed a co-culture model that enables the study of stromal cells with artificial capillaries using conventional imaging methods. We envision that acellular chips with two seeding ports can be readily shipped worldwide and could potentially be adopted as a new technology to study microvascular biology in a reproducible manner.
Gene editing (GE) pushes livestock breeding toward greater productivity and sustainability. Omics can help assess the environmental safety of gene-edited animals but comes with limitations. In this article, we propose a tiered framework for assessing how gene-edited livestock interact with ecosystems.
Organ fibrosis is a major cause of organ failure and mortality, yet effective disease-reversing therapeutics remain limited. Increasing evidence identifies BRD4, a BET family epigenetic reader, as a central regulator of fibrotic progression across organs. BRD4 integrates upstream injury signals, including TGFβ, NF-κB, oxidative stress, and mechanotransduction, to sustain transcription of pro-fibrotic and pro-inflammatory genes, promote myofibroblast activation, and reinforce pathological cellular plasticity. Across multiple organs and tissues, BRD4 contributes to extracellular matrix deposition, epithelial/endothelial-to-mesenchymal transition, inflammatory amplification, and fibrogenic cell-state maintenance. Preclinical studies show that pharmacological inhibition of BRD4 can attenuate or even reverse fibrosis across multiple organ systems. Emerging modalities, including BRD4 PROTACs, molecular glues, tissue-targeted degraders, microRNA-based modulation, and epigenome editing, further broaden the therapeutic potential of BRD4-directed strategies. Collectively, BRD4 represents a convergence node for chronic injury responses and a promising anti-fibrotic target. A deeper understanding of its context-specific functions, biomarker-guided targeting, and delivery strategies will be essential for translating BRD4-targeted interventions into effective therapies for fibrotic diseases.