Genome engineering with Cas9 and AAV repair templates, successes and pitfalls

Mammalian Genome, Год журнала: 2025, Номер unknown

Опубликована: Янв. 13, 2025

Genome editing, in particular the CRISPR/Cas9 system, is widely used to generate new animal models. However, generation of mutations, such as conditional knock-out or knock-in, can remain complex and inefficient, because difficulty deliver donor DNA (single double stranded) into nucleus fertilized oocytes. The use recombinant adeno-associated viruses (rAAV) a rapidly developing approach that promises improve efficiency creation In this mini-review, we explore progress challenges using combination with rAAV for precise genome editing. We will summarise current knowledge transduction, data on its rodent embryos easily sequence replacements insertions, limitations unexpected events observed date, protocol optimisations already place facilitate

Язык: Английский

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siRNA-based Delivery Systems: Technologies, Carriers, Applications, and Approved Products

European Journal of Pharmacology, Год журнала: 2025, Номер unknown, С. 177441 - 177441

Опубликована: Фев. 1, 2025

Язык: Английский

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Therapeutic potential of microRNA in meniscal repair and regeneration

The Knee, Год журнала: 2025, Номер 55, С. 18 - 23

Опубликована: Апрель 10, 2025

Язык: Английский

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Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles

Journal of Nanobiotechnology, Год журнала: 2025, Номер 23(1)

Опубликована: Фев. 20, 2025

Despite the outstanding clinical success of immunotherapy, its therapeutic efficacy in glioblastoma (GBM) is still limited. To identify critical regulators GBM immunity, we constructed a mouse single-guide RNA (sgRNA) library corresponding to all disease-related immune genes, and performed an vivo CRISPR knockout (KO) screen syngeneic models. We demonstrated that deletion GDF15 cells ameliorated immunosuppressive tumor microenvironment (TME) enhanced antitumor checkpoint blockade (ICB) response. Moreover, designed unique nanoparticles for efficient encapsulation CRISPR-Cas9, noninvasive brain delivery cell targeting, demonstrating effective safe strategy gene therapy. The CRISPR-Cas9 nanoparticles, known as ANPSS (Cas9/sgRNA), are easily created by enclosing single Cas9/sgRNA complex polymer shell sensitive glutathione. This also contains dual-action ligand aids crossing blood‒brain barrier, targeting cells, selectively releasing Cas9/sgRNA. Our encapsulating promising resulting high editing efficiency within tumors while showing minimal off-target high-risk tissues. Treatment with (Cas9/sgGDF15) effectively halted growth, reversed suppression, ICB These results emphasize potential role modulating enhancing effectiveness current immunotherapy strategies GBM. 1. In screens driver escape. 2. Synthesis TME-responsive 3. therapy enhances

Язык: Английский

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Molecular mechanisms of lncRNA NEAT1 in the pathogenesis of liver-related diseases, with special focus on therapeutic approaches

Pathology - Research and Practice, Год журнала: 2025, Номер 269, С. 155867 - 155867

Опубликована: Фев. 26, 2025

Язык: Английский

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Exploring the promise of lipoplexes: From concept to clinical applications

International Journal of Pharmaceutics, Год журнала: 2025, Номер 674, С. 125424 - 125424

Опубликована: Март 4, 2025

Язык: Английский

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Comparing the transfection efficiency of cationic monomer ratios in vinylimidazole and aminoethyl methacrylate copolymers

International Journal of Pharmaceutics X, Год журнала: 2025, Номер 9, С. 100327 - 100327

Опубликована: Март 4, 2025

Employing polycations as non-viral gene delivery vectors has been extensively studied owing to their safety, efficiency and possibility of modifying them in an intended way compared with viral vectors. However, the main challenge is finding a biocompatible transfection-efficient polymer. In this study, 2-aminoethyl methacrylate (A) 1-vinyl imidazole (V) were copolymerized at three different molar ratios by free radical polymerization method novel narrow molecular weight distribution obtained. The resulting copolymers used for condensation plasmid DNA (pDNA) N/P followed physicochemical characterizations polyplexes. At ratio 2, nanoplexes smaller than 120 nm. optimum formulations stable presence polyanions capable protecting condensed pDNA against nucleases. polyplexes having V A 1:1 most efficient carrier transfecting HeLa cells introduced promising vector.

Язык: Английский

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“ Advancing Nephrogenetics: Utilizing Single-Cell RNA Sequencing and Organoid Models for Precision Kidney Disease Treatment”

Current Pediatrics Reports, Год журнала: 2025, Номер 13(1)

Опубликована: Март 17, 2025

Язык: Английский

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CRISPR/Cas9-Based therapeutics as a promising strategy for management of Alzheimer’s disease: progress and prospects

Frontiers in Cellular Neuroscience, Год журнала: 2025, Номер 19

Опубликована: Апрель 7, 2025

CRISPR/Cas9 technology has revolutionized genetic and biomedical research in recent years. It enables editing modulation of gene function with an unparalleled precision effectiveness. Among the various applications prospects this technology, opportunities it offers unraveling molecular underpinnings a myriad central nervous system diseases, including neurodegenerative disorders, psychiatric conditions, developmental abnormalities, are unprecedented. In review, we highlight CRISPR/Cas9-based therapeutics as promising strategy for management Alzheimer’s disease transformative impact on AD research. Further, emphasize role generating accurate models identification novel therapeutic targets, besides CRISPR-based therapies aimed at correcting AD-associated mutations modulating processes. Furthermore, delivery systems reviewed potential non-viral nanotechnology-based carriers overcoming critical limitations effective is discussed. Overall, review highlights promise intricate processes underlying development AD, discusses its limitations, ethical concerns several challenges efficient across BBB, ensuring specificity, avoiding off-target effects. This article can be helpful better understanding based approaches way forward utilizing enormous targeted, gene-specific treatments that could change trajectory debilitating incurable illness.

Язык: Английский

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Developing a process of lentivirus purification from recombinant fluids using peptide affinity ligands

Bioengineering & Translational Medicine, Год журнала: 2025, Номер unknown

Опубликована: Апрель 7, 2025

Abstract Lentiviral vectors (LVVs) are emerging as an enabling tool in gene and cell therapies, yet the toolkit for purifying them at scale is still immature. A pivoting moment LVV isolation technology was marked by introduction of affinity ligands LVVs pseudo‐typed with Vesicular Stomatitis Virus G (VSV‐G) protein. Camelid antibody were initially discovered utilized to functionalize a resin capacity 10 14 particles per liter (vp/L). Shortly thereafter, our team introduced VSV‐G‐targeting peptides assessed their application from HEK293 harvests. In this study, we these develop novel resins and—first field—affinity membranes optimal binding capacity, productivity, removal host contaminants. To that end, evaluated different material, particle pore size, functional density, well fiber morphology, porosity, ligand distribution. The lead peptide‐functionalized membrane featured high (5 × 9 1.2 transducing units mL adsorbent, TU/mL) productivity (2.9 1.7 TU/mL min) afforded substantial enrichment cell‐transducing reduction contaminants (110–170‐fold) eluates. Finally, demonstrated purification process four steps: clarification nuclease treatment, capture bind‐and‐elute mode, polishing flow‐through ultra/dia‐filtration sterile filtration. processes yields 33%–46%, residual HCP level below 5 ng/mL, 1.25–1.5 active hour adsorbent.

Язык: Английский

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