Gene transfer into mammalian cells by particle bombardment.
Using COS-7 and Chinese hamster ovary cells as model systems, I have examined the efficiency of gene transfer into mammalian cells by particle bombardment. The most important parameters affecting transformation efficiency are the size of the particles, the target distance, and the extent of chamber vacuum. The size of the cell culture plate also affects transformation efficiency. Factors which have little effect on transformation efficiency are the helium pressure, the gap distance, and the macrocarrier travel distance. Compared to several other gene transfer techniques, particle bombardment has the advantage of requiring a low amount of DNA and a low number of cells for successful expression, measured as either transient or stable. I also describe transformation of several murine cell lines which have not been successfully transformed, or have been transformed at only low levels using other methods. These cell lines include preadipocytes (BMS-2), macrophages (J774), and transformed pre-B cells (38B9 and 70Z/3). Compared to transformation by electroporation, lipofection, and diethylaminoethyl dextran, particle bombardment was found to give 50- to 240-fold higher levels of transient expression as measured by luciferase activity in cell extracts.
Heiser
W C
WC
Genetic Systems Division, Bio-Rad Laboratories, Hercules, California 94547.
D000818:Animals; D016466:CHO Cells / Q000502:physiology; D002461:Cell Line, Transformed; D001517:Coleoptera / Q000201:enzymology; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D008156:Luciferases / Q000235:genetics; D008722:Methods; D051379:Mice; D008954:Models, Biological; D010316:Particle Size; D014170:Transformation, Genetic
D004247:DNA; D008156:Luciferases