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Transfection of a DNA/protein complex into nuclei of mammalian cells using polyoma capsids and electroporation
Authors:Wolf Bertling
Affiliation:(1) University of North Carolina at Chapel Hill, 27514 Chapel Hill, NC, USA
Abstract:We used empty capsids ofpolyoma virus to transfer DNA fragments and DNA/protein complexes into human cells. We encapsulated labeled and unlabeled single stranded DNA fragments by viral capsids. A complex of DNA with a DNA binding protein, recA, will also be taken up by the capsids, whereas the free protein is not incorporated. We further compared this gentle biological method of DNA transfection with a well-established physical method, electroporation. Electroporation also allows the transfer of DNA as well as protein into cells, although there is no proof that a DNA/protein complex can survive the procedure functionally. Whereas the viability of capsid transfected cells is unaffected (100%), electroporation reduces the viability to 90–95%. On the other hand, the amount of DNA found in the nucleus of electroporated cells is higher than for cells treated with loaded viral capsids.
Keywords:polyoma capsids  electroporation  DNA/protein transfection  lymphocytes
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