Efficient gene delivery and silencing of mouse and human pancreatic islets.
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.
Lefebvre
Bruno
B
Univ Lille Nord de France, F-59000 Lille, France. bruno.lefebvre@inserm.fr
Vandewalle
Brigitte
B
Longue
Justine
J
Moerman
Ericka
E
Lukowiak
Bruno
B
Gmyr
Valery
V
Maedler
Kathrin
K
Kerr-conte
Julie
J
Pattou
François
F
D000818:Animals; D017209:Apoptosis; D027682:Cation Transport Proteins / Q000235:genetics; D002470:Cell Survival; D002478:Cells, Cultured; D018274:Electroporation; D020868:Gene Silencing; D006801:Humans; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D000075703:Zinc Transporter 8
D027682:Cation Transport Proteins; D034741:RNA, Small Interfering; C516348:Slc30a8 protein, mouse; D000075703:Zinc Transporter 8