High efficiency lipid-based siRNA transfection of adipocytes in suspension.
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.


RESULTS
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
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.
Kilroy
Gail
G
Ubiquitin Biology Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA, USA.
Burk
David H
DH
Floyd
Z Elizabeth
ZE
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism*; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D008954:Models, Biological; D034741:RNA, Small Interfering / Q000378:metabolism*; D014162:Transfection / Q000295:instrumentation* / Q000379:methods*
D008055:Lipids; D034741:RNA, Small Interfering