high efficiency lipid-based sirna transfection of adipocytes in suspension小干扰rna转染效率高lipid-based悬挂的脂肪细胞.pdfVIP

High Efficiency Lipid-Based siRNA Transfection of Adipocytes in Suspension 1 2 1 Gail Kilroy , David H. Burk , Z. Elizabeth Floyd * 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 Abstract Background: Fully 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 Findings: We 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/Significance: Transfection 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. Citation: Kilroy G, Burk DH, Floyd ZE (2009) High Efficiency Lipid-Based siRNA Transfection of Adipocytes in Suspension. PLoS ONE 4(9): e6940. doi:10.1371/ journal.pone.0006940 Editor: Aimin Xu, University of Hong Kong, China Received May 21, 2009; Accepted August 7, 2009; Published September 11, 2009 Copyright: 2009 Kilroy et al. This is an open-access article distributed under the term