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Efficient gene delivery and silencing of mouse and human pancreatic islets

Authors :
Bruno Lukowiak
Valery Gmyr
Ericka Moerman
François Pattou
Kathrin Maedler
Brigitte Vandewalle
Julie Kerr-Conte
Justine Longue
Bruno Lefebvre
Source :
BMC Biotechnology, Vol 10, Iss 1, p 28 (2010), BMC Biotechnology
Publication Year :
2010
Publisher :
Springer Science and Business Media LLC, 2010.

Abstract

Background In view of the importance of beta cells in glucose homeostasis and the profound repercussions of beta cell pathology on human health, the acquisition of tools to study pancreatic islet function is essential for the design of alternative novel therapies for diabetes. One promising approach toward this goal involves the modification of gene expression profile of beta cells. Results This study describes a new method of gene and siRNA delivery into human pancreatic islets by microporation technology. We demonstrated that mild islet distention with accutase greatly enhanced the transfection efficiency without compromising in vitro function (secretion, apoptosis and viability). As an example, the recently identified gene involved in type 2 diabetes, ZnT8, can be over-expressed or silenced by RNA interference using this technology. Microporation can also be used on rodent islets. Conclusions Taken together, our results demonstrate that microporation technology can be used to modify gene expression in whole rodent and human islets without altering their in vitro function and will be key to the elucidation of the factors responsible for proper islet function.

Details

ISSN :
14726750
Volume :
10
Database :
OpenAIRE
Journal :
BMC Biotechnology
Accession number :
edsair.doi.dedup.....0fba9c043f512c5d0743ec632c9f24af