Biolistic and diolistic transfection: using the gene gun to deliver DNA and lipophilic dyes into mammalian cells.
Delivery of genes by firing them into mammalian cells is becoming increasingly popular for cells such as neurones, that have proved difficult to transfect by conventional means. In addition it is becoming apparent that this technique also provides a suitable method of introducing cell-specific dyes into mammalian tissues. Here we describe improved protocols for the rapid and efficient delivery of both DNA and lipophilic dyes into mammalian cells in both monolayers and in brain slices. The results show that transfection of genes, such as that for enhanced yellow fluorescent protein, using a modified microcarrier preparation technique combined with either a conventional or novel accelerator channel, yields rapid and efficient production of protein, which can be visualised in 24-48 h. Following similar delivery of lipophilic dyes neurones can be visualised in minutes. These techniques therefore provide an excellent means not only to examine the structure and function of neurones and neuronal genes, but also those of other cell types.
O'Brien
John
J
Neurobiology Division, MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
Lummis
Sarah C R
SC
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D019470:Biolistics / Q000295:instrumentation*; D001921:Brain / Q000648:ultrastructure; D002232:Carbocyanines / Q000378:metabolism; D002460:Cell Line; D004396:Coloring Agents / Q000378:metabolism*; D004247:DNA / Q000235:genetics*; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D009474:Neurons / Q000648:ultrastructure; D014162:Transfection / Q000379:methods*
D001426:Bacterial Proteins; D002232:Carbocyanines; D004396:Coloring Agents; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D004247:DNA