Sign in to use this feature.

Years

Between: -

Subjects

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Journals

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Article Types

Countries / Regions

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Search Results (4,359)

Search Parameters:
Keywords = Crispr

Order results
Result details
Results per page
Select all
Export citation of selected articles as:
45 pages, 3725 KB  
Review
Combating White Spot Syndrome Virus (WSSV) in Global Shrimp Farming: Unraveling Its Biology, Pathology, and Control Strategies
by Md. Iftehimul, Neaz A. Hasan, David Bass, Abul Bashar, Mohammad Mahfujul Haque and Morena Santi
Viruses 2025, 17(11), 1463; https://doi.org/10.3390/v17111463 (registering DOI) - 31 Oct 2025
Abstract
White Spot Syndrome Virus (WSSV) is one of the most devastating viral pathogens affecting shrimp, causing severe economic losses to the global farmed shrimp trade. The globalization of live shrimp trade and waterborne transmission have facilitated the rapid spread of WSSV across major [...] Read more.
White Spot Syndrome Virus (WSSV) is one of the most devastating viral pathogens affecting shrimp, causing severe economic losses to the global farmed shrimp trade. The globalization of live shrimp trade and waterborne transmission have facilitated the rapid spread of WSSV across major shrimp-producing countries since its initial emergence. The present review gives an updated account of WSSV biology, pathology, transmission dynamics, and recent developments in control measures. The virus, a double-stranded DNA virus of the Nimaviridae family, utilizes advanced immune evasion strategies, resulting in severe mortality. Shrimp lack adaptive immunity and hence rely predominantly on innate immunity, which is insufficient to mount an effective response against severe infections. Traditional disease control measures such as augmented biosecurity, selective breeding, and immunostimulants have, despite extensive research, achieved only limited success. New biotechnological tools such as RNA interference, CRISPR-Cas gene editing, and nanotechnology offer tremendous potential for disease mitigation. In parallel, the development of DNA and RNA vaccines targeting WSSV structural proteins, such as VP28, holds significant promise for stimulating the shrimp immune system. This review highlights the urgent need for a convergent approach to sustainable disease management in global shrimp aquaculture, with interdisciplinarity playing a pivotal role in shaping the future of WSSV control. Full article
(This article belongs to the Section Viral Immunology, Vaccines, and Antivirals)
Show Figures

Figure 1

20 pages, 950 KB  
Review
The Role of Plant Genetic Resources and Grain Variety Mixtures in Building Sustainable Agriculture in the Context of Climate Change
by Aleksandra Pietrusińska-Radzio, Paulina Bolc, Anna Tratwal and Dorota Dziubińska
Sustainability 2025, 17(21), 9737; https://doi.org/10.3390/su17219737 (registering DOI) - 31 Oct 2025
Abstract
In an era of global warming, sustainable agriculture, which emphasises the conservation of biodiversity and the rational use of natural resources, is growing in importance. One of the key elements is to increase the genetic diversity of crops through the use of crop [...] Read more.
In an era of global warming, sustainable agriculture, which emphasises the conservation of biodiversity and the rational use of natural resources, is growing in importance. One of the key elements is to increase the genetic diversity of crops through the use of crop wild relatives (CWRs) and local varieties, which provide a source of genes for resistance to biotic and abiotic stresses. Modern agricultural systems are characterised by low biodiversity, which increases the susceptibility of plants to diseases and pests. Growing mixtures of varieties, both intra- and interspecific, is a practical strategy to increase plant resistance, stabilise yields and reduce pathogen pressure. This manuscript has a review character and synthesises the current literature on the use of CWRs, local varieties, and variety mixtures in sustainable agriculture. The main research question of the study is to what extent plant genetic resources, including CWRs and local varieties, as well as the cultivation of variety mixtures, can promote plant resistance, stabilise yields and contribute to sustainable agriculture under climate change. The objectives of the study are to assess the role of genetic resources and variety mixtures in maintaining biodiversity and yield stability, and to analyse the potential of CWRs and local varieties in enhancing plant resistance. Additionally, the study investigates the impact of variety mixtures in reducing disease and pest development, and identifies barriers to the use of genetic resources in breeding along with strategies to overcome them. The study takes an interdisciplinary approach including literature and gene bank data analysis (in situ and ex situ), field trials of cultivar mixtures under different environmental conditions, genetic and molecular analysis of CWRs, the use of modern genome editing techniques (CRISPR/Cas9) and assessment of ecological mechanisms of mixed crops such as barrier effect, and induced resistance and complementarity. In addition, the study considers collaboration with participatory and evolutionary breeding programmes (EPBs/PPBs) to adapt local varieties to specific environmental conditions. The results of the study indicate that the integration of plant genetic resources with the practice of cultivating variety mixtures creates a synergistic model that enhances plant resilience and stabilises yields. This approach also promotes agroecosystem conservation, contributing to sustainable agriculture under climate change. Full article
27 pages, 2537 KB  
Article
NDR2 Kinase Regulates Microglial Metabolic Adaptation and Inflammatory Response: Critical Role in Glucose-Dependent Functional Plasticity
by Beatriz Fazendeiro, Ivo Machado, Anabela Rolo, Paulo Rodrigues Santos, António Francisco Ambrósio, Paulo F. Santos and Hélène Léger
Int. J. Mol. Sci. 2025, 26(21), 10630; https://doi.org/10.3390/ijms262110630 (registering DOI) - 31 Oct 2025
Abstract
Diabetic retinopathy (DR), a major complication of diabetes, is driven by chronic inflammation in which retinal microglial cells play a central role. The Hippo pathway kinases NDR1/2 regulate macrophage function, but their role in microglia and DR remain unknown. This study investigates the [...] Read more.
Diabetic retinopathy (DR), a major complication of diabetes, is driven by chronic inflammation in which retinal microglial cells play a central role. The Hippo pathway kinases NDR1/2 regulate macrophage function, but their role in microglia and DR remain unknown. This study investigates the function of the NDR2 kinase in microglial cells under high-glucose (HG) conditions. Using CRISPR-Cas9, we partially knocked out the Ndr2/Stk38l gene in BV-2 mouse microglial cells and analyzed metabolic activity, phagocytosis, migration, and cytokine release. We confirmed NDR2 expression in microglia and observed increased levels under HG, suggesting a role in hyperglycemia-induced stress. Ndr2/Stk38l (hereafter referred to as Ndr2) downregulation impaired mitochondrial respiration and reduced metabolic flexibility, indicating defective stress adaptation. Functionally, microglia with a partial downregulation of Ndr2 displayed reduced phagocytic and migratory capacity—both dependent on cytoskeletal dynamics. Moreover, Ndr2 downregulation altered the secretory profile, elevating pro-inflammatory cytokines (IL-6, TNF, IL-17, IL-12p70) even under normal glucose levels. These findings identify NDR2 protein kinase as a key regulator of microglial metabolism and inflammatory behavior under diabetic conditions. By modulating immune and metabolic responses, NDR2 may contribute to the neuroinflammatory processes underlying DR. Targeting NDR2 function in microglia may offer novel therapeutic strategies to mitigate retinal inflammation and progression of DR. Full article
(This article belongs to the Section Molecular Immunology)
Show Figures

Graphical abstract

41 pages, 3718 KB  
Article
Behavioral Balance in Tryptophan Turmoil: Regional Metabolic Rewiring in Kynurenine Aminotransferase II Knockout Mice
by Ágnes Szabó, Zsolt Galla, Eleonóra Spekker, Diána Martos, Mónika Szűcs, Annamária Fejes-Szabó, Ágnes Fehér, Keiko Takeda, Kinuyo Ozaki, Hiromi Inoue, Sayo Yamamoto, Péter Monostori, József Toldi, Etsuro Ono, László Vécsei and Masaru Tanaka
Cells 2025, 14(21), 1711; https://doi.org/10.3390/cells14211711 (registering DOI) - 31 Oct 2025
Abstract
Background: Cognitive, emotional, and social impairments are pervasive across neuropsychiatric conditions, where alterations in the tryptophan (Trp)–kynurenine pathway and its product kynurenic acid (KYNA) from kynurenine aminotransferases (KATs) have been linked to Alzheimer’s disease, Parkinson’s disease, depression, and post-traumatic stress disorder. In novel [...] Read more.
Background: Cognitive, emotional, and social impairments are pervasive across neuropsychiatric conditions, where alterations in the tryptophan (Trp)–kynurenine pathway and its product kynurenic acid (KYNA) from kynurenine aminotransferases (KATs) have been linked to Alzheimer’s disease, Parkinson’s disease, depression, and post-traumatic stress disorder. In novel CRISPR/Cas9-engineered KAT II knockout (aadat−/− also known as kat2−/−) mice, we observed despair-linked depression-like behavior with peripheral excitotoxicity and oxidative stress. KAT II’s role and its crosstalk with serotonin, indole-pyruvate, and tyrosine–dopamine remain unclear. It is unknown whether deficits extend to cognitive, emotional, motor, and social domains or whether brain tissues mirror peripheral stress. Objectives: Delineate domain-wide behaviors, brain oxidative/excitotoxic profiles, and pathway interactions attributable to KAT II. Results: Behavior was unchanged across strains. kat2−/− deletion remodeled Trp metabolic pathways: 3-hydroxykynurenine increased, xanthurenic acid decreased, KYNA fell in cortex and hippocampus but rose in striatum, quinaldic acid decreased in cerebellum and brainstem. These region-specific changes indicate metabolic stress across the brain and align with higher oxidative load and signs of excitotoxic pressure. Conclusions: Here, we show that KAT II deletion reshapes regional Trp metabolism and amplifies oxidative and excitotoxic imbalance. Although domain-wide behavioral measures, spanning cognition, sociability, and motor coordination, remained largely unchanged, these neurochemical alterations signify a latent emotional bias rather than overt depressive-like behavior. This work, therefore, refines prior findings by delineating KAT II–linked biochemical vulnerability as a potential substrate for stress-reactive affective dysregulation. Full article
28 pages, 3921 KB  
Review
Microdroplet Systems for Gene Transfer: From Fundamentals to Future Perspectives
by Mishell Criollo, Gina Layedra, Camilo Pérez-Sosa, Gustavo Rosero and Ana Belén Peñaherrera-Pazmiño
Micromachines 2025, 16(11), 1245; https://doi.org/10.3390/mi16111245 (registering DOI) - 31 Oct 2025
Abstract
Microfluidics enables precise control of fluid movement within microchannels, facilitating the generation of microdroplets at high frequencies. This technology provides a unique platform for conducting biological and chemical experiments, enhancing throughput and sensitivity, particularly in single-cell analysis. The microdroplet environment enhances interactions between [...] Read more.
Microfluidics enables precise control of fluid movement within microchannels, facilitating the generation of microdroplets at high frequencies. This technology provides a unique platform for conducting biological and chemical experiments, enhancing throughput and sensitivity, particularly in single-cell analysis. The microdroplet environment enhances interactions between cells and gene delivery materials, resulting in greater contact area, higher reagent concentration, and improved diffusion for both eukaryotic and prokaryotic cells. This review discusses the advantages and limitations of transfection and transformation within microdroplet technologies, highlighting their potential to improve gene editing efficiency while addressing challenges related to delivery mechanisms and cellular uptake rates. The integration of microdroplet technology with advanced gene editing tools, such as CRISPR/Cas9, promises to streamline processes and improve outcomes in various applications, including therapeutic interventions, vaccine development, regenerative medicine, and personalized medicine. These advancements could lead to more precise targeting of genetic modifications, resulting in tailored therapies that better meet individual patient needs. Overall, the integration of gene delivery in microdroplets represents a significant leap in biotechnology, enhancing the efficacy of gene delivery systems and opening new avenues for research and development in precision medicine. Full article
Show Figures

Figure 1

19 pages, 536 KB  
Review
Strengths and Limitations of Salicylic Acid Reporters
by Viktor V. Morozov, Ilia V. Yampolsky, Bulat K. Iskakov and Anastasia V. Balakireva
Int. J. Mol. Sci. 2025, 26(21), 10610; https://doi.org/10.3390/ijms262110610 (registering DOI) - 31 Oct 2025
Abstract
Salicylic acid (SA) is a key phytohormone that coordinates plant innate immunity and systemic acquired resistance. Because SA levels and signaling are highly dynamic in space and time, a suite of SA-focused tools, including SA-specific microbial biosensors and SA-responsive transcriptional and chemical reporters, [...] Read more.
Salicylic acid (SA) is a key phytohormone that coordinates plant innate immunity and systemic acquired resistance. Because SA levels and signaling are highly dynamic in space and time, a suite of SA-focused tools, including SA-specific microbial biosensors and SA-responsive transcriptional and chemical reporters, has been developed to study them. This review compares three classes of tools in terms of sensitivity, specificity, temporal resolution, invasiveness, quantifiability, and suitability across species. We describe developing genetically encoded sensors that can directly sense salicylic acid and report it, for example, via a fluorescence resonance energy transfer signal or another real-time output. We offer recommendations on method selection by research goal and plant species, as well as combined protocols (long-term autoluminescence plus local probes/biosensors) for cross-validation. Future work should prioritize substrate-free, quantitative SA reporters deployable in crops and the field; coupled with CRISPR-based editing and screening, these tools would enable reporter-guided discovery of immunity genes and rapid engineering of durable disease resistance. Full article
(This article belongs to the Topic Salicylic Acid as Plant Biostimulant)
Show Figures

Figure 1

26 pages, 1642 KB  
Review
Genome Editing Against HPV-Driven Cancers: From Bench to Clinic
by Muharrem Okan Cakir, Melis Selek, Betul Yilmaz, Mustafa Ozdogan and Gholam Hossein Ashrafi
Acta Microbiol. Hell. 2025, 70(4), 41; https://doi.org/10.3390/amh70040041 - 31 Oct 2025
Abstract
Genome editing technologies, including CRISPR/Cas9, TALENs, and ZFNs, offer promising approaches to disrupt HPV oncogenes E6 and E7, thereby restoring tumor-suppressor pathways. In this review, we summarize recent preclinical findings demonstrating selective apoptosis and tumor regression in HPV-positive cell and animal models, as [...] Read more.
Genome editing technologies, including CRISPR/Cas9, TALENs, and ZFNs, offer promising approaches to disrupt HPV oncogenes E6 and E7, thereby restoring tumor-suppressor pathways. In this review, we summarize recent preclinical findings demonstrating selective apoptosis and tumor regression in HPV-positive cell and animal models, as well as early-phase clinical studies exploring local CRISPR-based therapies. We also compare the relative strengths and limitations of major editing platforms, discuss delivery strategies, and highlight their potential integration with immunotherapy and conventional treatments. While preclinical studies show encouraging efficacy (e.g., up to 60% tumor regression in xenograft models and marked reactivation of p53/pRb pathways), translation into routine practice remains limited by challenges such as efficient delivery, minimizing off-target effects, long-term safety, cost, and ethical considerations. Continued optimization of high-fidelity nucleases, tissue-specific delivery systems, and genotype-tailored guide RNAs will be essential. Genome editing therefore represents a potential future addition to the therapeutic landscape of HPV-related diseases, but substantial barriers must be addressed before clinical implementation. Full article
Show Figures

Figure 1

47 pages, 1332 KB  
Review
Base and Prime Editing for Inherited Retinal Diseases: Delivery Platforms, Safety, Efficacy, and Translational Perspectives
by Haoliang Zhang, Yuxuan Li, Jiajie Li, Xiaosa Li and Tong Li
Pharmaceutics 2025, 17(11), 1405; https://doi.org/10.3390/pharmaceutics17111405 - 30 Oct 2025
Viewed by 14
Abstract
Inherited retinal diseases (IRDs) are a clinically and genetically heterogeneous spectrum of disorders that lead to progressive and irreversible vision loss. Gene therapy is the most promising emerging treatment for IRDs. While gene augmentation strategies have demonstrated clinical benefit and results within the [...] Read more.
Inherited retinal diseases (IRDs) are a clinically and genetically heterogeneous spectrum of disorders that lead to progressive and irreversible vision loss. Gene therapy is the most promising emerging treatment for IRDs. While gene augmentation strategies have demonstrated clinical benefit and results within the first approved ocular gene therapy, their application is restricted by adeno-associated virus (AAV) packaging capacity and limited efficacy for dominant mutations. Recent breakthroughs in precision genome editing, particularly base editing (BE) and prime editing (PE), have provided alternatives capable of directly correcting pathogenic variants. BE enables targeted single-nucleotide conversions, whereas PE further allows for precise insertions and deletions, both circumventing the double-strand DNA cleavage or repair processes typically induced by conventional CRISPR–Cas editing systems, thereby offering advantages in post-mitotic retinal cells. Preclinical investigations across murine and non-human primate models have demonstrated the feasibility, molecular accuracy, and preliminary safety profiles of these platforms in targeting IRD-associated mutations. However, critical challenges remain before clinical application can be realized, including limited editing efficiency in photoreceptors, interspecies variability in therapeutic response, potential risks of off-target effects, and barriers in large-scale vector manufacturing. Moreover, the delivery of genome editors to the outer retina remains suboptimal, prompting intensive efforts in capsid engineering and the development of non-viral delivery systems. This review synthesizes the current progress in BE and PE optimization, highlights innovations in delivery platforms that encompass viral and emerging non-viral systems and summarizes the major barriers to clinical translation. We further discuss AI-driven strategies for the rational design of BE/PE systems, thereby outlining their future potential and perspectives in the treatment of IRDs. Full article
(This article belongs to the Special Issue Ophthalmic Drug Delivery, 3rd Edition)
Show Figures

Graphical abstract

30 pages, 2754 KB  
Review
Recognition Element-Based Strategies for Rapid Detection of Foodborne Pathogens: Recent Progress and Perspectives
by Wang Guo, Meifeng Jiang, Yunkai Xie, Hong Xu and Zongbao Sun
Biosensors 2025, 15(11), 717; https://doi.org/10.3390/bios15110717 - 29 Oct 2025
Viewed by 121
Abstract
The detection of foodborne pathogens is of great significance for safeguarding food safety and public health. In recent years, rapid detection technologies based on diverse recognition elements have advanced considerably, driven by progress in molecular biology, materials science, and information technology. This review [...] Read more.
The detection of foodborne pathogens is of great significance for safeguarding food safety and public health. In recent years, rapid detection technologies based on diverse recognition elements have advanced considerably, driven by progress in molecular biology, materials science, and information technology. This review takes recognition elements as the central theme and systematically outlines the mechanisms and research progress of antibodies, nucleic acid aptamers, nucleic acid amplification techniques, CRISPR/Cas systems, molecular imprinting technology, peptides, and small-molecule receptors in foodborne pathogen detection, while comparing their performance in terms of specificity, sensitivity, stability, and applicability. In addition, this review further elaborates on the developmental trends of detection platforms, including multi-target and multimodal integration, microfluidics combined with portable point-of-care testing (POCT) systems, and intelligent terminals empowered by artificial intelligence algorithms. These trends provide new perspectives for improving detection systems in terms of throughput, portability, and intelligence. Overall, this review aims to serve as a comprehensive reference for the development of rapid, accurate, and intelligent detection systems for foodborne pathogens. Full article
(This article belongs to the Section Environmental Biosensors and Biosensing)
Show Figures

Figure 1

22 pages, 926 KB  
Review
Regulatory Mechanisms of Total Soluble Solids in Tomato: From QTL Mapping to Gene Editing
by Minghua Xu, Shujing Ji, Shengqun Pang, Yongen Lu, Shouming Li and Wei Xu
Foods 2025, 14(21), 3692; https://doi.org/10.3390/foods14213692 - 29 Oct 2025
Viewed by 226
Abstract
Total Soluble Solids (TSS) in tomatoes is a core indicator for evaluating fruit quality and processing characteristics. Its composition mainly consists of soluble sugars (such as fructose and glucose) and organic acids (such as citric acid and malic acid). The contents of sugars [...] Read more.
Total Soluble Solids (TSS) in tomatoes is a core indicator for evaluating fruit quality and processing characteristics. Its composition mainly consists of soluble sugars (such as fructose and glucose) and organic acids (such as citric acid and malic acid). The contents of sugars and acids and their ratio directly affect the flavor and nutritional value. Cultivated tomatoes have a TSS of 4–6%, compared with 10–15% in wild varieties. In recent years, with the advancement of molecular biology and genomics technologies, significant progress has been made in the research on the regulatory mechanisms of tomato fruit TSS and major sugars and acids, including the identification of major quantitative trait locus (QTLs) (Lin5, SlALMT9), functional characterization via CRISPR/Cas9 and elucidation of the transporter network. Breaking the negative correlation between TSS and yield remains a major bottleneck in breeding. Analyzing the mechanism by which environmental factors regulate the TSS and optimizing cultivation measures are crucial for increasing the TSS content in tomatoes. The deep integration of cutting-edge technologies (such as Genome-wide association studies (GWAS), metabolome-wide association studies (mGWAS), Genomic selection (GS), genome editing, and crop modeling) with design breeding is expected to accelerate the development of high-TSS tomato varieties. This paper reviews the current research status from the following four aspects: QTL mapping related to tomato TSS and mining of major genes, metabolic and transport mechanisms of major sugars and acids and key genes, the influence of environmental factors on TSS, and application of genetic improvement strategies and technologies. Full article
Show Figures

Figure 1

19 pages, 919 KB  
Review
CRISPR-Mediated Genome Editing in Peanuts: Unlocking Trait Improvement for a Sustainable Future
by Seong Ju Han, Jia Chae, Hye Jeong Kim, Jee Hye Kim, Young-Soo Chung, Sivabalan Karthik and Jae Bok Heo
Plants 2025, 14(21), 3302; https://doi.org/10.3390/plants14213302 - 29 Oct 2025
Viewed by 151
Abstract
Advancements in genome editing have transformed agricultural biotechnology by allowing for precise modifications of DNA. This technology has sparked increasing interest in enhancing important traits of major crops, including peanuts. As a nutritionally rich legume prized for its high oil content, peanut production [...] Read more.
Advancements in genome editing have transformed agricultural biotechnology by allowing for precise modifications of DNA. This technology has sparked increasing interest in enhancing important traits of major crops, including peanuts. As a nutritionally rich legume prized for its high oil content, peanut production still faces significant challenges, including disease outbreaks, nutrient deficiencies, and pest infestations. Addressing these challenges is essential for achieving high yields and sustainable cultivation. CRISPR technology, a cutting-edge genome editing tool, has emerged as a powerful platform for improving peanut traits. Its ability to facilitate gene knockouts, regulate gene expression, and introduce targeted genetic changes has accelerated research efforts in this field. The successful applications of CRISPR in peanut improvement, such as increasing oleic acid content and reducing allergenicity, reassure us about the effectiveness and potential of this technology. Despite the complexity of the peanut genome as a polyploid crop, these successes demonstrate the power of genome editing. This review emphasizes the crucial role of genome editing in enhancing peanut traits and outlines the promising future of CRISPR-based approaches in advancing peanut breeding and agricultural productivity. Full article
(This article belongs to the Special Issue Plant Transformation and Genome Editing)
Show Figures

Figure 1

19 pages, 4656 KB  
Article
Improving Gene Knock-In Efficiencies in Sheep Primary Cells Using a CRISPR/Cas9-Gal4 System
by Yan Li, Sujun Wu, Yunpeng Wu, Yiming Yuan, Yue Zhao, Teng Meng, Wensheng Zhang, Jin Wang and Yefeng Qiu
Curr. Issues Mol. Biol. 2025, 47(11), 899; https://doi.org/10.3390/cimb47110899 - 29 Oct 2025
Viewed by 178
Abstract
Currently, a major challenge exists in CRISPR-mediated genome editing research in sheep: the low efficiency of exogenous large DNA fragment targeted integration without drug selection or fluorescence enrichment. This restriction significantly impedes the use of precise genome editing in sheep for agricultural, biological, [...] Read more.
Currently, a major challenge exists in CRISPR-mediated genome editing research in sheep: the low efficiency of exogenous large DNA fragment targeted integration without drug selection or fluorescence enrichment. This restriction significantly impedes the use of precise genome editing in sheep for agricultural, biological, and biomedical purposes. In this study, we employed the strategy of increasing the local concentration of the homologous repair template at the site of the DNA double-strand break (DSB). We achieved this by fusing the DNA binding domain (BD) of the Gal4 protein (Gal4-BD) to the N-terminal end of the SpCas9 protein using a 32-amino acid (aa) flexible linker. Additionally, we incorporated a 17 bp UAS at the 3′ end of the donor template, which can be specifically recognized and bound by Gal-BD. As a result, we observed a significant improvement in the knock-in efficiency of the exogenous large DNA fragment (2997 bp) in sheep fetal fibroblasts (SFFs), increasing it from 5.30% (8/151) to 16.67% (32/192), providing a comparatively efficient and user-friendly method to promote CRISPR-mediated gene knock-in in sheep. Full article
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
Show Figures

Figure 1

21 pages, 3952 KB  
Article
Creating an Improved Diatoxanthin Production Line by Knocking Out CpSRP54 in the zep3 Background in the Marine Diatom Phaeodactylum tricornutum
by Charlotte Volpe, Zdenka Bartosova, Ralph Kissen, Per Winge and Marianne Nymark
Mar. Drugs 2025, 23(11), 419; https://doi.org/10.3390/md23110419 - 29 Oct 2025
Viewed by 253
Abstract
Diatoxanthin is a photoprotective carotenoid found in a few groups of microalgae displaying in vitro anti-inflammatory and anti-cancer properties, making it a promising candidate for nutraceutical, pharmaceutical, and cosmetic applications. However, large-scale production is currently nonexistent because of two major challenges: Instability during [...] Read more.
Diatoxanthin is a photoprotective carotenoid found in a few groups of microalgae displaying in vitro anti-inflammatory and anti-cancer properties, making it a promising candidate for nutraceutical, pharmaceutical, and cosmetic applications. However, large-scale production is currently nonexistent because of two major challenges: Instability during microalgae harvesting, where diatoxanthin is rapidly converted back to its inactive precursor diadinoxanthin under non-stressful light conditions, and dependence on prolonged exposure to high-intensity light, which is costly and technically challenging during indoor high-cell-density cultivation. The first limitation was previously addressed by knocking out zeaxanthin epoxidase 3 (ZEP3) in the marine diatom Phaeodactylum tricornutum, resulting in a mutant that stabilized diatoxanthin under non-stressful light conditions. Here, we report an improved diatoxanthin production line where both of the described challenges have been overcome. This was achieved by creating P. tricornutum mutants where the phenotype of the zep3 mutant was combined with the light-sensitive phenotype of the chloroplast signal recognition particle 54 (cpsrp54) mutant. Growth rates were maintained at wild-type levels at light intensities ≤ 150 µmol photons m−2 s−1 in the zep3cpsrp54 mutants, but prolonged medium light exposure resulted in a 1.5- and 7-fold increase in diatoxanthin concentration compared with zep3 and wild-type, respectively. When returned to low light, the zep3cpsrp54 cultures retained ~80% of their accumulated diatoxanthin. The improved production lines allow for diatoxanthin accumulation without the use of high-intensity light and with limited loss of diatoxanthin when returned to non-stressful light conditions. Full article
(This article belongs to the Special Issue High-Value Algae Products, 2nd Edition)
Show Figures

Figure 1

14 pages, 2853 KB  
Article
The Chromosome 19 microRNA Cluster Facilitates Cancer Stemness in Hepatocellular Carcinoma
by Marian T. Underwood, Varsha Devarapalli, Goodwin G. Jinesh, John H. Lockhart, Marco Napoli, Nino Mtchedlidze, Elsa R. Flores and Andrew S. Brohl
Non-Coding RNA 2025, 11(6), 74; https://doi.org/10.3390/ncrna11060074 - 29 Oct 2025
Viewed by 279
Abstract
Background/Objectives: Hepatocellular carcinoma (HCC) is one of the world’s deadliest cancers; however, the mechanisms that contribute to its aggressiveness are poorly understood. In the recent literature, overexpression of the Chromosome 19 MicroRNA Cluster (C19MC) has been associated with an aggressive phenotype and unfavorable [...] Read more.
Background/Objectives: Hepatocellular carcinoma (HCC) is one of the world’s deadliest cancers; however, the mechanisms that contribute to its aggressiveness are poorly understood. In the recent literature, overexpression of the Chromosome 19 MicroRNA Cluster (C19MC) has been associated with an aggressive phenotype and unfavorable prognosis in HCC. However, the molecular consequences of C19MC overexpression in HCC remain poorly understood. Methods: Here, we created a constitutive C19MC-overexpressing HCC model and used two different CRISPR-engineered C19MC-overexpressing HCC models to analyze phenotype and transcriptomic changes. Results: We observed that C19MC overexpression induces cancer stem cell (CSC) phenotypic features in vitro and analyzed transcriptomic changes in genes correlating with stemness, such as NFκB and EMT. Conclusions: C19MC induces changes in HCC that are consistent with stemness and aggression, which provides a better understanding of why C19MC could be a biomarker of poor prognosis. Full article
Show Figures

Figure 1

29 pages, 2575 KB  
Review
Technologies in Biomarker Discovery for Animal Diseases: Mechanisms, Classification, and Diagnostic Applications
by Salwa Eman, Raza Mohai Ud Din, Muhammad Hammad Zafar, Mengke Zhang, Xin Wen, Jiayu Ma, Ahmed A. Saleh, Hosameldeen Mohamed Husien, Mengzhi Wang and Xiaodong Guo
Animals 2025, 15(21), 3132; https://doi.org/10.3390/ani15213132 - 29 Oct 2025
Viewed by 361
Abstract
Animal diseases remain a major constraint to livestock productivity and public health, necessitating accurate, early diagnostic methods. This review examines the classification and mechanisms of diagnostic, prognostic, and predictive biomarkers in veterinary medicine and evaluates how advanced technologies enable their discovery. Mechanistically, biomarkers [...] Read more.
Animal diseases remain a major constraint to livestock productivity and public health, necessitating accurate, early diagnostic methods. This review examines the classification and mechanisms of diagnostic, prognostic, and predictive biomarkers in veterinary medicine and evaluates how advanced technologies enable their discovery. Mechanistically, biomarkers function as molecular indicators of disease presence, progression, or therapeutic response, and are essential in species where clinical signs often appear late or are non-specific. We detail the contribution of high-throughput omics platforms, genomics (NGS, RNA-Seq), proteomics (LC-MS/MS, DIGE), and metabolomics (NMR, LC-MS/MS) in identifying disease-specific molecular signatures. Emerging technologies, including CRISPR/Cas9, AI-enhanced imaging, aptamer-based biosensors, and microfluidic devices, show significant diagnostic potential. Case studies, including canine melanoma, bovine respiratory disease complex (BRDC), and congenital portosystemic shunts in dogs, illustrate the real-world applicability of biomarkers. Challenges such as a lack of standardization, species variability, and poor clinical translation are acknowledged. The review concludes that integrating biomarker mechanisms with advanced analytical technologies is key to advancing veterinary diagnostics and disease control. Full article
Show Figures

Figure 1

Back to TopTop