Gene Ther
Division of Biochemistry, School of Biological Sciences, Royal Holloway-University of London, Egham, Surrey, UK.
Published: October 2001
We have examined the ability of proliferating myoblasts and post-mitotic, differentiated myotubes to produce retroviral vector using hybrid adeno-retroviral vectors as templates. We show that production of retroviral vector from myoblasts peaks 48 h after adenoviral infection at 4.8 x 10(4) cfu/ml and is scarcely detectable by 96 h. Both fully and partially differentiated myotubes were able to generate a sustained increase in the levels of retroviral vector compared with myoblasts peaking 48 h at 1.4 x 10(5) cfu/ml and 1.8 x 10(5) cfu/ml, respectively. Addition of the cell cycle inhibitor aphidicolin (5 microg/ml) had no effect on the production of retroviral vector from fully differentiated myotubes, but resulted in an 80% increase in vector production from partially differentiated myotubes. Thus indicating that retroviral vector production is more efficient in post-mitotic myotubes and is independent of muscle cell cycle progression.
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Sci Rep
March 2025
Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger-Center of Molecular Medicine (NFZ), Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
The pBMN-I-GFP plasmid is a widely used retroviral vector for producing retroviral particles, utilized by thousands of laboratories worldwide. However, we observed that E. coli transformed with pBMN-I-GFP failed to grow on selective LB agar plates containing ampicillin or carbenicillin, in contrast to similar retroviral vectors.
View Article and Find Full Text PDFMethods Mol Biol
March 2025
Department of Biochemistry and Molecular Biology, Pharmacy Faculty, Complutense University of Madrid, Madrid, Spain.
Development of the programmable nuclease Ca9 has increased our understanding of the molecular basis of the physiological processes and diseases demonstrating a remarkable potential across various fields, including basic research, applied biotechnology, and biomedical research. CRISPR-Cas9 is a gene-editing technique that uses a guide RNA to direct the Cas9 enzyme to specific DNA sequences, where it creates double-strand breaks. These breaks can be repaired through either nonhomologous end joining (NHEJ), leading to gene mutations, or homology-directed repair (HDR), enabling precise edits.
View Article and Find Full Text PDFMethods Mol Biol
March 2025
Univ Lyon, Université Claude Bernard Lyon 1, INSERM, Stem Cell and Brain Research Institute U1208, Bron, France.
Lineage reprogramming of glial cells into induced neurons (iNs) has emerged as an innovative strategy to replace neurons lost due to injury or neurological diseases. Here, we describe a step-by-step protocol to induce in vivo conversion of reactive glial cells, proliferating within the injured hippocampus, into mature and functional GABAergic iNs through retrovirus-mediated expression of two neurogenic fate determinants (Ascl1 and Dlx2). We have previously applied this method to study the integration and functional impact of GABAergic iNs in epileptic mice (Lentini et al.
View Article and Find Full Text PDFNat Commun
March 2025
San Raffaele Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Liver fibrosis occurs in several genetic and acquired disease conditions, leading to alterations of the tissue and metabolism, which may adversely affect viral vector-mediated gene therapy. Here, we assessed the impact of liver fibrosis on in vivo gene transfer to hepatocytes mediated by lentiviral vectors or adeno-associated viral vectors. We exploited two chemically induced fibrosis mouse models characterized by tissue damage in different areas of the liver lobule.
View Article and Find Full Text PDFAnn Allergy Asthma Immunol
March 2025
Division of Immunology, Allergy and Retrovirology, Department of Pediatrics. Baylor College of Medicine. Houston and The Woodlands, TX. Electronic address:
Hematopoietic stem cell transplant has been the single curative treatment for inborn errors of immunity (IEI) and is recommended for the most severe IEI conditions, such as severe combined immunodeficiency (SCID). However, adverse outcomes primarily due to histocompatibility differences between the donor and the patient are still of concern. Progress in genetic and molecular mechanisms, as well as new technology to insert DNA sequences in cell genomes has allowed the development of strategies to treat genetic diseases by correcting the gene defect in patients' cells.
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