Journal
DEVELOPMENTAL DYNAMICS
Volume 235, Issue 8, Pages 2210-2219Publisher
WILEY
DOI: 10.1002/dvdy.20873
Keywords
lipefection; transfection; pegylation; cationic lipids
Categories
Funding
- NICHD NIH HHS [5T32 HD07491] Funding Source: Medline
- NIDCD NIH HHS [DC04185] Funding Source: Medline
- NIDDK NIH HHS [DK066445] Funding Source: Medline
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We have used cationic lipid-based transfection reagents for ectopic gene expression experiments in developing vertebrate embryos. Lipofectamine, Lipofectamine 2000, and Lipofectamine enhanced with a disulfide linked pegylated lipid (mPEG-SS-DOPE) were initially tested and optimized in cell culture. Two reagent formulations, 1:4 (DNA:Lipofectamine 2000) Lipofectamine 2000, and 7.5% pegylated Lipofectamine, produced the highest levels of gene expression in vitro. Those formulations, containing the enhance green fluorescent protein reporter gene, were microinjected into intact vertebrate embryos-systemically through the vasculature and locally into selected tissues-to assess in vivo transfection efficiency. Whereas both formulations are capable of transfecting cells in developing embryos in vivo, greater transfection efficiencies in a broader range of tissue types were obtained with the pegylated Lipofectamine formulation. We conclude that in developing vertebrate embryos, optimized cationic lipid-based reagents are capable of producing significant levels of ectopic gene expression and can be used as alternatives to electroporation and viral-mediated gene delivery.
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