Single-cell electroporation for gene transfer in vivo.
We report an electroporation technique for targeting gene transfer to individual cells in intact tissue. Electrical stimulation through a micropipette filled with DNA or other macromolecules electroporates a single cell at the tip of the micropipette. Electroporation of a plasmid encoding enhanced green fluorescent protein (GFP) into the brain of intact Xenopus tadpoles or rat hippocampal slices resulted in GFP expression in single neurons and glia. In vivo imaging showed morphologies, dendritic arbor dynamics, and growth rates characteristic of healthy cells. Coelectroporation of two plasmids resulted in expression of both proteins, while electroporation of fluorescent dextrans allowed direct visualization of transfer of molecules into cells. This technique will allow unprecedented spatial and temporal control over gene delivery and protein expression.
Haas
K
K
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Sin
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Javaherian
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Li
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Cline
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D000818:Animals; D046508:Culture Techniques; D003911:Dextrans / Q000008:administration & dosage; D018274:Electroporation; D019793:Fluorescein / Q000008:administration & dosage; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000378:metabolism; D007395:Interneurons / Q000378:metabolism; D007814:Larva / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal; D009474:Neurons / Q000737:chemistry / Q000378:metabolism; D017966:Pyramidal Cells / Q000378:metabolism; D051381:Rats; D012235:Rhodamines / Q000008:administration & dosage; D013477:Superior Colliculi / Q000737:chemistry / Q000378:metabolism; D014162:Transfection / Q000379:methods; D014981:Xenopus
D003911:Dextrans; D008164:Luminescent Proteins; D012235:Rhodamines; D049452:Green Fluorescent Proteins; C005358:tetramethylrhodamine; D019793:Fluorescein