tailieunhanh - Báo cáo khoa học: " Targeting lentiviral vector to specific cell types through surface displayed single chain antibody and fusogenic molecule"

Targeting lentiviral vector to specific cell types through surface displayed single chain antibody and fusogenic molecule | Lei et al. Virology Journal 2010 7 35 http content 7 1 35 VIROLOGY JOURNAL RESEARCH Open Access Targeting lentiviral vector to specific cell types through surface displayed single chain antibody and fusogenic molecule Yuning Lei Kye-Il Joo Jonathan Zarzar Clement Wong Pin Wang Abstract Background Viral delivery remains one of the most commonly used techniques today in the field of gene therapy. However one of the remaining hurdles is the off-targeting effect of viral delivery. To overcome this obstacle we recently developed a method to incorporate an antibody and a fusogenic molecule FM as two distinct molecules into the lentiviral surface. In this report we expand this strategy to utilize a single chain antibody SCAb for targeted transduction. Results Two versions of the SCAb were generated to pair with our various engineered FMs by linking the heavy chain and the light chain variable domains of the anti-CD20 antibody aCD20 via a GS linker and fusing them to the hinge-CH2-CH3 region of human IgG. The resulting protein was fused to either a HLA-A2 transmembrane domain or a VSVG transmembrane domain for anchoring purpose. Lentiviral vectors generated with either version of the SCAb and a selected FM were then characterized for binding and fusion activities in CD20-expressing cells. Conclusion Certain combinations of the SCAb with various FMs could result in an increase in viral transduction. This two-molecule lentiviral vector system design allows for parallel optimization of the SCAb and FMs to improve targeted gene delivery. Introduction Gene therapy is the introduction of a functional gene into a dysfunctional cell for a therapeutic benefit. To date viral vectors remain the most commonly used gene delivery vehicles due to their high transduction efficiencies 1 2 . In particular lentiviral vectors represent one of the most effective gene delivery vehicles as they allow for stable long-term transgene expression in both dividing and non-dividing .