A novel, membrane receptor-based retroviral vector for Fanconi anemia group C gene therapy

Gene Therapy
A W MachlManfred Kubbies

Abstract

Retroviral vectors are effective shuttle systems by introducing therapeutically relevant genes stably into the genome of proliferating cells. The majority of vectors applied for research or clinical applications use neomycin for cell selection and identification. To circumvent the time consuming and potentially toxic G418 selection process in transduction studies we constructed a novel marker vector using I-NGFR as a cell surface marker to identify DNA repair defective Fanconi anemia cells complemented with the FAC gene. The new vector constructed is based on a MoMLV backbone, a signal peptide-deleted I-NGFR receptor gene under control of a LTR promoter and the therapeutically relevant FAC gene placed downstream of a SV40 promoter. Supernatants containing high titers of amphotropic viruses from FACS cloned cell cultures were obtained and tested for primary transduction rates, rapid detection of transduced cells within 48 h and correction of mitomycin C-induced cell cycle G2 phase accumulation in a single assay using multiparameter, dual laser flow cytometry. Primary transduction efficiency detected via (I-NGFR) antibody was between 5% and 30% with Fanconi cell lines, 5% with CD34+ cells and 15% with PBLs. MMC-induced G2 phase c...Continue Reading

Citations

Mar 29, 2002·Journal of Cellular Physiology·Thorsten HoevelManfred Kubbies
Dec 4, 2003·International Journal of Cancer. Journal International Du Cancer·Thorsten HoevelManfred Kubbies
Jan 18, 2003·World Journal of Gastroenterology : WJG·Che ChenJie-Shou Li

❮ Previous
Next ❯

Related Concepts

Related Feeds

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.