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The emergence of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome-editing system has brought about a significant revolution in the realm of managing human diseases, establishing animal models, and so on. To fully harness the potential of this potent gene-editing tool, ensuring efficient and secure delivery to the target site is paramount. Consequently, developing effective delivery methods for the CRISPR/Cas9 system has become a critical area of research. In this review, we present a comprehensive outline of delivery strategies and discuss their biomedical applications in the CRISPR/Cas9 system. We also provide an in-depth analysis of physical, viral vector, and non-viral vector delivery strategies, including plasmid-, mRNA- and protein-based approach. In addition, we illustrate the biomedical applications of the CRISPR/Cas9 system. This review highlights the key factors affecting the delivery process and the current challenges facing the CRISPR/Cas9 system, while also delineating future directions and prospects that could inspire innovative delivery strategies. This review aims to provide new insights and ideas for advancing CRISPR/Cas9-based delivery strategies and to facilitate breakthroughs in biomedical research and therapeutic applications.
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http://dx.doi.org/10.1016/j.ajps.2023.100854 | DOI Listing |
Curr Gene Ther
September 2025
Centre for Research Impact & Outcome-Chitkara College of Pharmacy, Chitkara University, Rajpura, Punjab 140401, India.
Gene therapy has revolutionized the therapeutic landscape for hemophilia A and B, offering the prospect for persistent endogenous production of coagulation factors VIII and IX. Recent advances in adeno-associated virus (AAV)-mediated gene transfer, particularly the approvals of valoctocogene roxaparvovec (Roctavian) and etranacogene dezaparvovec (Hemgenix), mark significant milestones in hemophilia care. This mini-review synthesizes emerging clinical data from phase I-III trials published between 2022 and 2025, emphasizing efficacy, durability, and immunogenicity profiles of leading AAV-based therapies.
View Article and Find Full Text PDFClin Chim Acta
September 2025
Department of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical, Tehran, Iran. Electronic address:
Acute myeloid leukemia (AML) represents a genetically heterogeneous malignancy, with mutations in the nucleophosmin-1 (NPM1) gene identified as the most prevalent and clinically significant molecular biomarkers. These mutations play a crucial pivotal role in the realms of diagnosis, prognosis, and therapeutic decision-making. Although an ideal measurable residual disease (MRD) test has yet to be developed, there is increasing acknowledgment of the significance of advanced molecular methodologies for monitoring MRD in NPM1-mutated (NPM1) AML.
View Article and Find Full Text PDFAgeing Res Rev
September 2025
Department of Pharmaceutical Toxicology, School of Pharmacy, China Medical University, Shenyang, 110122, China; Laboratory of Naturel Medicine for drug discovery, School of Pharmacy, China Medical University, Shenyang, 110122, China. Electronic address:
Calcium (Ca)/calmodulin (CaM)-dependent protein kinase II (CaMKII) is an emerging drug target for age-related diseases. It is a multifunctional kinase with complex activation modes, numerous isoforms, broad tissue distribution, and a dual role in health and disease. In particular, its isoforms share a high degree of conservation within the catalytic and regulatory domains, with only minor differences confined to the linker region.
View Article and Find Full Text PDFAdv Sci (Weinh)
September 2025
Shanghai Key Laboratory of Maternal Fetal Medicine, Shanghai Institute of Maternal-Fetal Medicine and Gynecologic Oncology, Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University, Shanghai, 201204, China.
Syncytiotrophoblasts (STBs) constitute one of the core components of the placenta, responsible for synthesizing pregnancy-sustaining hormones such as human chorionic gonadotropin (HCG). Deficient syncytialization of cytotrophoblasts affects the hormonal secretion and placental development, contributing to pregnancy-associated disorders, including spontaneous miscarriage. To date, the molecular mechanisms, particularly the role of transcription factors (TFs), in STB lineage specification remain incompletely understood.
View Article and Find Full Text PDFMicroPubl Biol
August 2025
Faculty of Science, Yamagata University.
In yeast, mitochondrial fission is mediated by the dynamin-like GTPase Dnm1, which is recruited to the mitochondrial outer membrane by its receptor, Fis1. To investigate the spatial distribution of Fis1, we used the CRISPR-Cas9 system to insert the gene fragment encoding mNeonGreen into the gene for its N-terminal tagging. Fluorescence microscopy revealed that mNeonGreen-Fis1 appeared as discrete puncta on mitochondria, in addition to a diffuse signal.
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