Development of high-concentration lipoplexes for in vivo gene function studies in vertebrate embryos
Article first published online: 29 JUL 2011
Copyright © 2011 Wiley-Liss, Inc.
Volume 240, Issue 9, pages 2108–2119, September 2011
How to Cite
Geetha-Loganathan, P., Nimmagadda, S., Hafez, I., Fu, K., Cullis, P. R. and Richman, J. M. (2011), Development of high-concentration lipoplexes for in vivo gene function studies in vertebrate embryos. Dev. Dyn., 240: 2108–2119. doi: 10.1002/dvdy.22708
- Issue published online: 15 AUG 2011
- Article first published online: 29 JUL 2011
- Manuscript Accepted: 3 JUL 2011
- Canadian Institutes of Health Research (CIHR). Grant Numbers: MOP49431, MOP 77524
- chicken embryo;
- mouse embryo;
- cationic lipids;
- interference RNA;
- micromass culture
Here we report that highly concentrated cationic lipid/helper lipid-nucleic acid complexes (lipoplexes) can facilitate reproducible delivery of a variety of oligonucleotides and plasmids to chicken embryos or to mouse embryonic mesenchyme. Specifically, liposomes composed of N,N-dioleyl-N,N-dimethylammonium chloride (DODAC)/1,2 dioleoyl glycero-3-phosphorylethanolamine (DOPE) prepared at 18-mM concentrations produced high levels of transfection of exogenous genes in vivo and in vitro. Furthermore, we report sufficient uptake of plasmids expressing interference RNA to decrease expression of both exogenous and endogenous genes. The simplicity of preparation, implementation, and relatively low toxicity of this transfection reagent make it an attractive alternative for developmental studies in post-gastrulation vertebrate embryos. Developmental Dynamics 240:2108–2119, 2011. © 2011 Wiley-Liss, Inc.