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Efficient gene delivery and silencing of mouse and human pancreatic islets
- 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.
- Subjects :
- Cell Survival
lcsh:Biotechnology
Apoptosis
Zinc Transporter 8
Gene delivery
Biology
Methodology article
Transfection
Mice
Insulin-Secreting Cells
lcsh:TP248.13-248.65
medicine
Animals
Humans
Glucose homeostasis
Gene silencing
Pancreatic islet function
Gene Silencing
RNA, Small Interfering
Cation Transport Proteins
Cells, Cultured
Microscopy, Confocal
Reverse Transcriptase Polymerase Chain Reaction
Pancreatic islets
Molecular biology
Mice, Inbred C57BL
Electroporation
medicine.anatomical_structure
Cancer research
Stem cell
Beta cell
Biotechnology
Subjects
Details
- ISSN :
- 14726750
- Volume :
- 10
- Database :
- OpenAIRE
- Journal :
- BMC Biotechnology
- Accession number :
- edsair.doi.dedup.....0fba9c043f512c5d0743ec632c9f24af