Search Results (4,132)

Bacterial sexually transmitted and sexually associated infections remain a major global health concern, increasingly complicated by antimicrobial resistance and the limited effectiveness of existing therapies. In this context, bacteriophage-based and phage-derived approaches have re-emerged as potential alternative antibacterial strategies. This narrative review examines their applicability across key bacterial pathogens associated with sexually transmitted infections, including Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Treponema pallidum and biofilm-associated bacterial vaginosis, with a particular focus on pathogen-specific biological barriers. Available evidence indicates that the success of phage-based interventions is strongly dependent on factors such as intracellular localisation, structural characteristics of the bacterial envelope and the presence of polymicrobial biofilms. While phage-derived platforms, including endolysins, depolymerases and engineered phages, demonstrate antibacterial activity in experimental settings, their effectiveness is uneven across different pathogens. Biofilm-associated infections appear more accessible to these approaches, whereas intracellular and structurally atypical bacteria are currently considered more challenging targets based on available mechanistic and experimental evidence. These observations highlight the need for pathogen-specific engineering strategies and delivery systems. Overall, phage-based therapeutics in this field should be considered within a framework that integrates biological constraints with targeted antimicrobial design. Full article

Glycosylation is essential for hematopoietic cell homeostasis and malignant transformation. Dysregulated expression of glycosylation genes in leukemia cells accelerates disease progression and fosters drug resistance. Therefore, targeting these genes offers a promising avenue for anti-leukemic therapy. In this study, we explore the roles of N-glycans in acute promyelocytic leukemia (APL) differentiation using the ATRA-induced wild-type NB4 (WT/ATRA) or HL-60 cell model. We found that expression of N-acetylglucosaminyltransferase IVa (GnT-IVa, encoded by the MGAT4A gene) and its product (β1,4-GlcNAc-branched N-glycan) increased significantly during differentiation, as evaluated by lectin blot, real-time PCR, and flow cytometry. Interestingly, analysis of the Gene Expression Omnibus (GEO) public data showed that MGAT4A expression is significantly lower in APL patients, and higher MGAT4A expression was associated with favorable survival in AML cohorts. To address the role of GnT-IVa in differentiation, we established MGAT4A- and MGAT4B-knockout (KO) NB4 cell lines using CRISPR/Cas9. Compared to WT/ATRA cells, MGAT4A KO, but not MGAT4B KO, markedly suppressed ATRA-induced differentiation, as evidenced by reduced expression of CD11b and CD11c. We found that CD11b is a major glycoprotein carrying β1,4-GlcNAc-branched N-glycans. This modification enhanced CD11b stability, as CD11b expression declined more rapidly in MGAT4A KO cells in the presence of cycloheximide. In addition, MGAT4A KO suppressed ERK/MAPK signaling, which contributed to differentiation. Our study highlights the critical role of GnT-IVa in regulating APL differentiation, which may provide a basis for developing new differentiation therapies for APL. Full article

Graphitic carbon nitride (g-C₃N₄), an emerging metal-free semiconductor material, has attracted considerable attention in the field of photoelectrochemical (PEC) sensing due to its unique electronic structure, excellent chemical stability, and visible-light responsiveness. This article systematically reviews recent advances in research on g-C₃N₄-based PEC sensors applied to water environment monitoring. First, the fundamental physicochemical properties of g-C₃N₄ are introduced, along with its advantages and limitations in PEC sensing applications. Subsequently, four main performance enhancement strategies are outlined: heterojunction construction (including type II, Z-scheme, and S-scheme heterojunction), elemental doping and defect engineering, morphology control and nanostructure design, as well as various signal amplification approaches such as self-powered systems, dual-mode detection, and cyclic amplification. Furthermore, the current application status of these sensors in detecting typical water pollutants, including heavy metal ions (e.g., Pb²⁺, Cu²⁺, Cd²⁺, Hg²⁺), antibiotics (e.g., tobramycin, norfloxacin, kanamycin), pesticide residues (e.g., chlorpyrifos, atrazine, glyphosate), and pathogenic microorganisms (e.g., Salmonella, Candida albicans), is comprehensively reviewed, with particular emphasis on detection sensitivity, selectivity, and real-sample performance. Finally, the remaining challenges in terms of long-term stability, anti-interference capabilities in complex matrices, portability, and multifunctional integration are analyzed, and future development directions are proposed, including smartphone-based intelligent sensing, CRISPR/Cas12a-assisted signal amplification, and multi-target high-throughput detection. This review aims to provide a reference for the rational design and practical application of g-C₃N₄-based PEC sensors in the field of water environment monitoring. Full article

Anthracnose, caused by Colletotrichum gloeosporioides, is a globally distributed phytopathogenic disease with a broad host range, posing a serious threat to the healthy growth of forest trees, including Cunninghamia lanceolata. To enable rapid and accurate on-site detection of this pathogen, this study developed a comprehensive field-deployable detection method. The approach integrates the EZ-D method (EASY DNA extraction) for rapid nucleic acid extraction with recombinase polymerase amplification (RPA) and the CRISPR/Cas12a system. A specific target gene, designated Cglo6922, was identified for the detection of C. gloeosporioides. The entire detection process can be completed within approximately 25 min, comprising a 10-min isothermal RPA at 39 °C followed by a 15-min Cas12a cleavage reaction. Specificity evaluation showed that the method successfully detected two C. gloeosporioides isolates derived from different hosts, while no cross-reactivity was observed against a panel of 32 other isolates, including ten Colletotrichum species, eight Phytophthora species, six Pythium species, seven Fusarium species, and one Botryosphaeria dothidea isolate, demonstrating robust species-level specificity. Sensitivity testing revealed that the method achieved a limit of detection (LOD) of 10 pg/μL of genomic DNA for C. gloeosporioides. Furthermore, by incorporating the EZ-D rapid extraction method (requiring only one minute for DNA extraction at a cost of approximately $0.03 USD per sample), target nucleic acid was successfully extracted from artificially inoculated Cunninghamia lanceolata branch samples and proved compatible with the RPA-CRISPR/Cas12a detection system. In conclusion, this study establishes a novel field-deployable detection method for C. gloeosporioides that is rapid, cost-effective, highly specific, and highly sensitive, providing a powerful tool for point-of-care testing (POCT) of this disease. Full article

Synthetic biology is reshaping in vitro diagnostics (IVD) by enabling programmable and modular biosensing elements that can be integrated into point-of-care testing (POCT) platforms. Compared with conventional assays that depend on fixed chemistries and centralized instrumentation, synthetic biology-based systems offer adaptable molecular recognition, tunable signal processing, and flexible readout formats for decentralized diagnostics. In this review, we present synthetic biology-enabled IVD as programmable biosensing platforms organized into four functional layers: molecular recognition, signal transduction and amplification, output generation, and system integration. We discuss four major enabling modules, including cell-free protein synthesis (CFPS) systems, aptamer and riboswitch sensors, CRISPR-Cas diagnostic platforms, and microfluidic integration technologies. We summarize representative clinical applications from 2021 to 2025 in infectious disease detection, cancer biomarker analysis, and drug metabolism/toxicity screening. In addition, we examine practical considerations beyond analytical sensitivity, including matrix tolerance, workflow complexity, manufacturability, quantitative capability, and regulatory readiness. Finally, we highlight future directions for programmable diagnostics, including AI-assisted biosensor design, multimodal readouts, interoperable platform architectures, and real-world clinical validation. Full article

Sickle cell disease (SCD) is a monogenic disorder responsible for recurrent vaso-occlusive crises, progressive organ damage, and shortened life expectancy. For decades, allogeneic hematopoietic stem cell transplantation from a matched sibling donor has been the only established cure, but its reach remains limited by donor availability and transplant-related toxicity. The approval of two autologous gene therapy products in 2023, exagamglogene autotemcel (exa-cel) and lovotibeglogene autotemcel (lovo-cel), marked a turning point for the SCD population and the gene therapy field in general. This review proposes a molecular rationale for fetal hemoglobin reactivation and β-globin gene addition, describes the engineering of lentiviral and CRISPR-based platforms, and highlights the clinical evidence accumulated to date that demonstrated durable disease modification with acceptable short-term toxicity. We then assess the clinical positioning of gene therapy within the broader spectrum of curative options compared to current available treatments and address the financial, ethical and psychosocial barriers that limit access to gene therapy both within high-income countries and globally. Critical research priorities include long-term safety surveillance, comparative effectiveness studies, pediatric trials below 12 years, and validated patient-reported outcome instruments. Base editing, non-genotoxic conditioning, and in vivo delivery represent the most promising avenues to broaden access and reduce treatment burden. Full article

The innate immune system, particularly the retinoic acid-inducible gene I (RIG-I)-like receptor (RLR) signaling pathway, is a major early defense barrier against influenza A virus infection. However, excessive immune responses can trigger lethal cytokine storms and severe immune-mediated pathology. In this study, we performed a genome-wide CRISPR/dCas9 gene activation screen in human lung epithelial (A549) cells by using an A/Puerto Rico/8/1934 (H1N1) reporter virus, and identified the ubiquitin-specific protease USP17L13 as a novel negative regulator of innate immunity that promotes influenza virus replication. Overexpression of USP17L13 significantly enhanced the replication of multiple subtypes of influenza viruses in A549 cells, including a human pandemic H1N1 virus, seasonal H3N2 viruses, as well as a globally circulating clade, 2.3.4.4b, of the highly pathogenic avian H5N1 virus. Transcriptomic analysis demonstrated that USP17L13 suppresses host antiviral defenses by downregulating nuclear factor kappa B (NF-κB) signaling and arachidonic acid metabolism, while upregulating pathways associated with ribosomal translation and oxidative phosphorylation to facilitate viral production. Mechanistically, USP17L13 attenuates the host interferon (IFN) response by promoting the degradation of the key viral RNA sensors, RIG-I, and melanoma differentiation-associated protein 5 (MDA5). Further analysis revealed that USP17L13 is inducible by type I and type II interferons as well as inflammatory cytokines, suggesting that it may act as a negative-feedback regulator to limit excessive inflammation. Collectively, our findings identify USP17L13 as a previously unrecognized proviral host factor and provide new insight into how host deubiquitinases shape influenza virus-host interactions, with potential implications for host-directed approaches to controlling excessive inflammation during viral infection and improving influenza vaccine production. Full article

Huanglongbing (HLB), primarily caused by ‘Candidatus Liberibacter asiaticus’ (CLas), threatens global citrus production. Deciphering the molecular interplay between citrus and CLas is crucial for successful control. This review synthesizes current understanding of the molecular mechanisms underlying citrus-CLas interactions, providing a comprehensive overview that spans immune signaling, hormonal and metabolic reprogramming, non-coding RNA-mediated regulation, pathogen effector biology, and emerging biotechnological interventions. We detail the hierarchical host response: initial immune recognition via pattern recognition receptors, triggering reactive oxygen species bursts and calcium signaling. Moreover, hormonal network reprogramming and their complex interplay in defense/susceptibility are examined. Transcriptomic studies have revealed key features of metabolic reprogramming, including suppression of photosynthesis and impairment of phloem function. Additionally, long-term strategies like cell wall reinforcement, accumulation of defensive compounds such as flavonoids and terpenoids, and roles of post-transcriptional regulation of microRNAs are discussed. Conversely, CLas counter-defense, notably effector-mediated immunity suppression and host metabolism manipulation, is also considered. Comparative transcriptomics between tolerant and susceptible varieties identifies tolerance or resistance genes/pathways for breeding and engineering. Despite this progress, critical knowledge gaps remain, particularly regarding the precise molecular mechanisms of CLas immune evasion and effector-mediated suppression, the genetic basis of natural tolerance, and the field-level efficacy of defense priming strategies. Future research directions should integrate single-cell omics, CRISPR/Cas9 editing, nano-enabled delivery, and microbiome engineering to bridge these gaps and accelerate HLB-tolerant/resistant citrus development. This review synthesizes how molecular profiling advances understanding of citrus defense mechanisms against HLB, and underscores the imperative for interdisciplinary research and global collaboration. Full article

The family of Nek kinases has 11 human members that are conserved in their kinase domains but diverse in their regulatory domains. Functionally, they can be associated with diverse aspects of cell cycle regulation, from mitosis and primary cilia function to centrosome disjunction in the G2 phase and checkpoints of the DNA damage response. However, novel functional contexts have emerged in recent years, including regulatory roles of Neks 1, 4, 5, and 10 in mitochondrial metabolic and morphological homeostasis. We recently generated, by CRISPR-Cas9 technology, a DU-145 prostate cancer cell line, with an NEK6 gene knockout. Here, we focus on a detailed characterization of changes in this cell line, in mitochondrial respiration function and morphology. DU-145 NEK6 knockout cells exhibited reduced mitochondrial respiration and a fragmented phenotype in electron microscopy, with reduced mitochondrial cristae numbers. Alterations in mitochondrial architecture and respiration were correlated with increased expression of anaerobic glycolytic proteins (HK2, PFKP, and LDHA) and decreased expression of PDH, an enzyme of aerobic glycolysis. Molecular analysis by Western blot revealed decreased levels of mitochondrial mass and biogenesis protein markers (TOM20, TFAM), without alterations in other markers such as VDAC1/3 or mtDNA copy number in the NEK6 knockout cells. Furthermore, the regulators of mitochondrial fusion/fission are altered in the knockout cells (decrease in the Long-OPA1:Short-OPA1 ratio and DRP1 total level), which is associated with an increase in endoplasmic reticulum–mitochondria contact at ≤20 nm observed in transmission electron microscopy (TEM) image analysis. Using analysis of TEM micrographs, we found an increase in the autophagic structures (autophagosome, amphisome, and autolysosome), with mitochondria as cargo in some structures, which was correlated with a decrease in LC3A/B and an increase in the BECLIN1 total level, and with an increase in acidic vesicles approximation, suggesting that reduction in TOM20 and TFAM without alterations in VDAC1/3 and mtDNA copy number might be related to mitochondrial degradation through autophagy. Together, our data suggest a new role for NEK6 in regulating mitochondrial homeostasis, where its loss alters mitochondrial morphology and respiration, and could be associated with an increase in the degradation of the dysfunctional mitochondria through autophagy. Full article

Background/Objectives: Rice fragrance is mainly determined by 2-acetyl-1-pyrroline (2-AP), which is negatively regulated by OsBADH2. However, the contribution of its paralog OsBADH1 to aroma-associated metabolism and GABA-linked abiotic stress responses remains unclear. This study investigated whether simultaneous disruption of OsBADH1 and OsBADH2 further enhances 2-AP accumulation while affecting stress tolerance in rice. Methods: Independent osbadh1 and osbadh2 knockout lines were generated using CRISPR/Cas9 and crossed to obtain homozygous osbadh1 osbadh2 double mutants. Wild type, single mutants, and double mutants were compared for 2-AP accumulation, GABA content, agronomic traits, abiotic stress responses, and expression of genes associated with GABA metabolism and stress responses. Results: The osbadh2 mutant showed a marked increase in 2-AP, and the osbadh1 osbadh2 double mutant exhibited the highest level, corresponding to a 7.1-fold increase over the wild type. In contrast, the GABA content progressively decreased and reached 0.46-fold of the wild-type level in the double mutant. Under normal growth conditions, the double mutant showed no major agronomic defects. However, under salinity and drought stress, its survival declined to 0.41-fold and 0.40-fold of the wild-type levels, respectively. KEGG and expression analyses further indicated coordinated disruption of GABA-associated metabolic and stress-responsive pathways in the double mutant. Conclusions: Combined disruption of OsBADH1 and OsBADH2 enhanced aroma-associated metabolism but weakened GABA-linked abiotic stress tolerance, revealing a trade-off between increased fragrance and reduced stress resilience in rice. Full article

Schistosomiasis, caused by Schistosoma species, is notoriously difficult to accurately diagnose with conventional methods. In this study, we present an innovative biosensor that integrates CRISPR–Cas12a technology with nucleic acid aptamers for the highly sensitive detection of Schistosoma japonicum eggs. The biosensor leverages a Y-shaped DNA structure (Y-DNA) that incorporates an aptamer specific to S. japonicum eggs, along with an activator DNA and a segment for immobilization on magnetic nanomaterials. Upon target recognition, the Y-DNA releases the activator, which triggers the collateral cleavage activity of Cas12a, enabling the direct detection of eggs. This system demonstrates remarkable sensitivity, being capable of detecting individual eggs in infected rabbit serum and feces. Moreover, it effectively distinguishes the eggs of S. japonicum from those of other parasitic species. The simplicity, high sensitivity, and rapid detection of our biosensor offer significant potential for improving the diagnosis of schistosomiasis, providing a novel, reliable tool for early detection in clinical settings. Full article

Gene editing technology, a revolutionary tool in molecular biology, enables precise modifications of genomic sequences and gene expression patterns, thereby conferring desired traits to cells or organisms. Since 2014, CRISPR/Cas9 has rapidly become the most widely used gene editing method in agricultural animals due to its high editing efficiency. Subsequently, the development of novel gene editing systems, such as base editors and prime editors, has provided enhanced precision and reduced off-target effects. These advancements have facilitated the transition of gene editing from laboratory research to clinical and agricultural applications. Gene editing has been extensively utilized to enhance production traits, improve disease resistance, facilitate disease detection, and establish disease models. This review outlines the development of gene editing technologies, discusses the advantages and limitations of key gene editing tools, and explores their applications in poultry. Furthermore, it examines the challenges and future prospects of gene editing in animal husbandry, including off-target effects, ethical concerns, and technical complexities. Full article

Helicobacter pylori (H. pylori) infection is closely associated with the development of chronic gastritis, peptic ulcers, and gastric cancer, highlighting the importance of rapid and accurate detection for disease prevention and clinical management. In this study, a one-pot LAMP-CRISPR/Cas12b assay targeting the CagA gene was developed for H. pylori detection. First, the LAMP system was optimized by systematically screening key reaction components. Subsequently, a one-step LAMP-CRISPR/Cas12b detection platform was established through optimization of the ratio between the LAMP premix and CRISPR buffer, reaction temperature, Cas12b concentration, and ssDNA reporter concentration. Under optimal conditions, the assay achieved a detection limit of 3.14 × 10¹ copies/µL, representing a tenfold improvement in sensitivity compared with conventional LAMP and PCR assays (3.14 × 10² copies/µL). In addition, the entire detection process could be completed within 1 h. Validation using 17 culture-positive and 17 culture-negative samples demonstrated complete concordance with culture-based results, with no false-positive or false-negative detections observed. These findings indicate that the proposed platform possesses high sensitivity, excellent specificity, rapid turnaround, and operational simplicity, demonstrating strong potential for point-of-care testing and applications in resource-limited settings. Full article

Protein turnover is essential for cellular metabolism, organelle biogenesis, stress adaptation, and ultimately the viability of cells and tissues. Papain-like cysteine proteases (PLCPs) are one of the vital components in protein degradation. PLCPs have been reported to act in senescence-associated proteolysis, but their roles in vegetative growth remain unclear. We identified PtCP1, an AALP-like PLCP in Populus tomentosa, localized to the vacuole and acid-triggered activated. CRISPR/Cas9-generated loss-of-function mutant (d7) showed dwarfism and non-stomatal photosynthetic limitations. On the other hand, the gain-of-function line (EM, deleted ERFNIN domain) exhibited accelerated growth and enhanced photosynthetic parameters. We showed d7 had the accumulation of Rubisco, which was the most important protein in photosynthetic carbon fixation. Transcriptomics revealed dysregulated carbon metabolism in d7. This data supported PtCP1-mediated proteolysis regulated photosynthetic carbon assimilation via altered Rubisco turnover, and then it increased the biomass accumulation during vegetative growth in woody plants. Full article

Tomato brown rugose fruit virus (ToBRFV) poses a major threat to global tomato (Solanum lycopersicum) production, as it can overcome conventional resistance genes that are effective against tobamoviruses. In this study, a multiplex CRISPR/Cas9 system was developed to target the SlTOM1 susceptibility gene family (SlTOM1a–d), which encodes host factors essential for tobamovirus replication. Six guide RNAs (gRNAs), designed following 12 off-target analyses, were assembled into a multiplex CRISPR/Cas9 construct using a Golden Gate cloning strategy and introduced into tomato genotypes through an Agrobacterium-based tissue culture transformation procedure. Although primary T₀ transformants exhibited chimeric mutation patterns, stable inheritance and segregation of edited alleles were confirmed in the T₁ generation. Sequence analyses identified diverse indel mutations across target loci, with SlTOM1d exhibiting the highest editing efficiency. Multiplex genome editing successfully generated single-, double-, and triple-mutant combinations, with higher-order mutants displaying the strongest tolerance phenotypes. Following mechanical ToBRFV inoculation, edited T₁ plants exhibited markedly reduced symptom severity, low viral accumulation, and improved fruit health compared to wild-type controls. RT-qPCR analysis further confirmed significantly reduced viral RNA levels, supporting a host-factor-mediated tolerance mechanism. Importantly, edited lines maintained normal growth and agronomic performance. Collectively, these findings demonstrate that multiplex CRISPR/Cas9-mediated targeting of SlTOM1 homologs represents a promising and practical strategy for improving ToBRFV tolerance in tomato breeding programs. Full article