Quantitative Comparison of Intracellular Trafficking and Nuclear Transcription between Adenoviral and Lipoplex Systems

摘要: To develop nonviral gene vectors that are sufficient for clinical application, it is necessary to understand why and what extent inferior viral vectors, which in general show a more efficient transfection activity. This study describes systematic quantitative comparison of the cellular uptake subsequent intracellular distribution (e.g., endosome/lysosome, cytosol, nucleus) exogenous DNA transfected by living cells, using combination TaqMan PCR recently developed confocal image-assisted three-dimensionally integrated quantification method. As model, adenovirus (Ad) Lipofectamine Plus (LFN) were used since they highly potent widely respectively. The findings indicate efficiency LFN significantly higher than Ad. Once taken up cell, Ad exhibited comparable endosomal escape slightly nuclear transfer compared with LFN. In contrast, requires 3 orders magnitude intranuclear copies exhibit transgene expression Ad, suggesting difference principally arises from differences transcription not trafficking between