High Efficiency Lipid-Based siRNA Transfection of Adipocytes in Suspension |
| |
Authors: | Gail Kilroy David H. Burk Z. Elizabeth Floyd |
| |
Affiliation: | 1. Ubiquitin Biology Laboratory, Pennington Biomedical Research Center, Baton Rouge, Louisiana, United States of America.; 2. Cell Biology and Bioimaging Core, Pennington Biomedical Research Center, Baton Rouge, Louisiana, United States of America.;University of Hong Kong, China |
| |
Abstract: | BackgroundFully differentiated adipocytes are considered to be refractory to introduction of siRNA via lipid-based transfection. However, large scale siRNA-based loss-of-function screening of adipocytes using either electroporation or virally-mediated transfection approaches can be prohibitively complex and expensive.Methodology/Principal FindingsWe present a method for introducing small interfering RNA (siRNA) into differentiated 3T3-L1 adipocytes and primary human adipocytes using an approach based on forming the siRNA/cell complex with the adipocytes in suspension rather than as an adherent monolayer, a variation of “reverse transfection”.Conclusions/SignificanceTransfection of adipocytes with siRNA by this method is economical, highly efficient, has a simple workflow, and allows standardization of the ratio of siRNA/cell number, making this approach well-suited for high-throughput screening of fully differentiated adipocytes. |
| |
Keywords: | |
|
|