Caecum: A Potential Site for Studying Gene Transfer in vivo

Abstract

Over the past years we have designed and synthesized a new class of synthetic vectors called phosphonolipids and then shown their ability to transfect lungs cells of mice with efficiency. One of them, GLB73, gave high levels of transfection. In the study reported here, we explored the potential of caecum as an alternative site for studying the feasibility of gene transfer in this site. Transfections were performed by using two reporter genes encoding for (igalactosidase and luciferase; transfection activity was assessed using two tests: chemiluminescent and cytofiuorimetric assays. The results obtained showed successful gene transfer into the caecum: up to 27% cells were LacZ+ with a mean of 11%; the maximum of efficiency was also observed 3 days after transfection which then decreased until day 7. Our lipoplex was 8-fold more efficient than the naked DNA (Mann Whitney test; p = 0.03). Moreover, we were able to visually follow the uptake of lipoplexes by enterocytes from 30 mn to 3 days post transfection.

So, this study constituted an encouraging first step in the assessment of the caecum as a potential model for gene transfer. In the near future, further electrophysiological studies using the gene of interest as CFTR gene should be performed in the caecum.

Keywords::

• Transfection
• Cationic-Lipids
• Caecum
• Model