stable transfectant - is there a way to make stables without antibiotic resistance? (May/25/2006 )
I need to make stable transfectants with a plasmid that doesn't express an antibiotic resistance for selection of eukariotic cells. Is there a way to select those stable transfectants?
Recombinantion is a rare event and usually has negative consequence toward cell growth. I guess thats why selection is usually needed. However, years ago one postdoc in our lab was on a project trying to find out if certain viral sequences would help insert his construct (contianing beta-gal reporter) to mammalian cells without forceful situation like antibiotic selection. He was albe to carry out the experiment for several months using limited seeding density and fluorecent substrate and hand pick colonies followed by expansion. Nowadays FACS or immunoselection can make it simpler. I know someone has done repeated FACS over time in K562 cells transfected with EGFP plasmid without selection pressure before. You can try a Pubmed search see if you can find that work.
Couldn't you lox out the antiobiotic resistant cassette after selection to prevent it from interfering with your results?
There are lots of ways to do this...