Transfection of a DNA/protein complex into nuclei of mammalian cells using polyoma capsids and electroporation.
We used empty capsids of polyoma 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.
Bertling
W
W
University of North Carolina, Chapel Hill 27514.
D002213:Capsid / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000378:metabolism*; D004560:Electricity; D005822:Genetic Vectors; D006801:Humans; D011120:Polyomavirus; D011938:Rec A Recombinases / Q000378:metabolism; D014162:Transfection
D004277:DNA, Single-Stranded; D003854:Deoxyribonucleotides; D004247:DNA; D011938:Rec A Recombinases