Enhanced DNA transfer into fish bone cells using polyethylenimine.
The use of in vitro cell culture systems to assess gene function largely depends on the successful transfer of DNA into target cells. Well developed in mammals, transfection methods are still to be optimized for non-mammalian cell culture systems, like fish. Here we describe a rapid, cost-efficient, and successful method to transfer DNA into a fish bone-derived cell line using polyethylenimine (PEI) as the DNA carrier. Using this method, DNA transfer was remarkably enhanced in comparison with commercially available reagents, as demonstrated by the increased activity of both luciferase and green fluorescent protein observed in the transfected cells. Its efficiency in transferring DNA into a wide range of cell types, including non-mammalian and hard-to-transfect cells, in addition to a low cost, show that PEI is a reagent of choice for nonviral vector transfection.
Braga
Daniel
D
Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal.
Laizé
Vincent
V
Tiago
Daniel M
DM
Cancela
M Leonor
ML
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D004337:Drug Carriers / Q000737:chemistry; D008422:Materials Testing; D010006:Osteoblasts / Q000378:metabolism*; D011094:Polyethyleneimine / Q000737:chemistry*; D021541:Sea Bream / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D020099:Coated Materials, Biocompatible; D004337:Drug Carriers; D011094:Polyethyleneimine; D004247:DNA