Cationic lipid-mediated gene transfer to the growing murine and human airway

Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways

“…naked plasmid DNA or DNA/liposomes) often lead to transient gene expression when delivered to the mouse lung, despite the persistence of intact plasmid DNA. [40][41][42] Strong viral promoters such as the cytomegalovirus (CMV IE) promoter can become attenuated in the lung, resulting in silencing of transgene expression after only a few days. 41,43 However, as we report here, CMVmediated reporter gene expression from rAAV5/5 is persistent under similar conditions.…”

Detection of plasmid DNA vectors following gene transfer to the murine airways

“…10 However, as with other plasmid DNA/liposome formulations, gene expression from GL67/plasmid DNA complexes is transient, lasting only a few days after administration. 15 Naked plasmid DNA has also been evaluated as a potential lung gene transfer vector and in spite of low levels of reporter gene expression 12 correction of the chloride defect in the nasal epithelium of CF patients was as effective as GL67/plasmid DNA. 16 Improved duration of gene expression was observed when viral promoters were replaced by endogenously expressed human promoters, such as elongation factor 1a or polyubiquitin B or C, resulting in reporter gene expression for up to 6 months post-administration to the murine lung.…”

Optimisation of real-time quantitative RT-PCR for the evaluation of non-viral mediated gene transfer to the airways

“…Results from other studies have determined that a large proportion of transgene expression following instillation of DNA/liposomes is located in the lung parenchyma, rather than in the conducting airways where CFTR expression is likely to be required. 29,30 The presence of mRNA does not necessarily indicate a comparable level of functional protein. In a clinical study evaluating DNA/liposome gene transfer to the nasal epithelium in CF subjects, 12 greater than 5% of the level of endogenous CFTR mRNA, was expressed, yet the CF chloride conductance defect was only partially corrected.…”