How do lentiviral vectors work

Large-scale collaborative efforts are underway to use lentiviruses to block the expression of a specific gene using RNA interference technology in high-throughput formats.

The expression of short-hairpin RNA shRNA reduces the expression of a specific gene , thus allowing researchers to examine the necessity and effects of a given gene in a model system. These studies can be a precursor to the development of novel drugs which aim to block a gene-product to treat a disease.

Another common application is to use a lentivirus to introduce a new gene into human or animal cells. For example, a model of mouse hemophillia is corrected by expressing wild-type platlet-factor VIII, the gene that is mutated in human hemophillia. Lentiviral infection have advantages over other gene therapy methods including high-efficiency infection of dividing and non-dividing cells, long-term stable expression of a transgene, and low immunogenicity.

Lentiviruses have also been successfully used for transfection of diabetic mice with the gene encoding PDGF platelet-derived growth factor , a therapy being considered for use in humans. These treatments, like most current gene therapy experiments, show promise but are yet to be established as safe and effective in controlled human studies. Lentiviral vectors: their molecular design, safety, and use in laboratory and preclinical research. Recent advances in lentiviral vector development and applications.

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Hum Gene Ther Clin Dev. Lentiviral-mediated phenotypic correction of cystic fibrosis pigs. JCI Insight. A third-generation lentivirus vector with a conditional packaging system. J Virol. Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery.

Biosafety Considerations for Research with Lentiviral Vectors. Available from: osp. Available from: www. Generation of high-titer pseudotyped retroviral vectors with very broad host range. Methods Cell Biol. Reversing a model of Parkinson's disease with in situ converted nigral neurons. Search-and-replace genome editing without double-strand breaks or donor DNA.

Biallelic RIPK1 mutations in humans cause severe immunodeficiency, arthritis, and intestinal inflammation. Nat Microbiol. Lentiviral Plasmids- Frequently Asked Questions. Neutrophil extracellular traps target senescent vasculature for tissue remodeling in retinopathy.

Enhanced CAR-T cell activity against solid tumors by vaccine boosting through the chimeric receptor. Somatic APP gene recombination in Alzheimer's disease and normal neurons. A compact synthetic pathway rewires cancer signaling to therapeutic effector release. The glycan CA promotes pancreatitis and pancreatic cancer in mice. EGR1 is a gatekeeper of inflammatory enhancers in human macrophages.

Sci Adv. Neutrophil extracellular traps produced during inflammation awaken dormant cancer cells in mice. Mol Cell. Nat Cell Biol. Charged polymers modulate retrovirus transduction via membrane charge neutralization and virus aggregation. Biophys J. Polybrene increases retrovirus gene transfer efficiency by enhancing receptor-independent virus adsorption on target cell membranes.

Biophys Chem. Mol Ther Methods Clin Dev. Improving viability and transfection efficiency with human umbilical cord wharton's jelly cells through use of a ROCK inhibitor.

Cell Reprogram. The significance of controlled conditions in lentiviral vector titration and in the use of multiplicity of infection MOI for predicting gene transfer events. To the same tube, 2. Polybrene 3 enhances the transduction efficiency by neutralizing the charges between viral particles and the cell membrane. However, the exact mechanism is not clearly known. Mix gently and plate the cells in a new 10cm dish.

Wait for 24 hours before changing to fresh complete medium without virus. If the shRNA construct contained a fluorescent marker, you can check the success of the integration 48 hours post-transfection; if not, go ahead with the antibiotic selection for another 48 hours. I usually start the transduction on a Wednesday, so the cells can be selected over the weekend. You Did It! Please feel free to comment below with any questions.

Thanks for reading and good luck! Morgan, and M. Polybrene increases retrovirus gene transfer efficiency by enhancing receptor-independent virus adsorption on target cell membranes. Facebook Twitter LinkedIn More. Written by Ramya Parimi. Image Credit: C. Thank you. Log in to Reply. Leave a Comment Cancel Reply You must be logged in to post a comment. Share via. Copy Link. Powered by Social Snap. Copy link. Copy Copied.