I encounter a problem when I try to establish a stable endothelial cell line (Ea.hy926 cells) expressing siRNA.
I transfectd cells with the vectors expressing target or negative control siRNA using electroporation and start the selection with G418 (200 ug/ml). I did find clones in cells transfected with target or negative control siRNA expression vector. But when I transferred the clones into 48 well plate and cultured them, even the negative control cells stopped growing and gradually died!!!
Does anyone have the experence with siRNA expression vectors? Thanks!
vector: pSilencer 3.1-H1 neo vector provided by Ambion
negative control siRNA template: also provided by Ambion
stable transfection with siRNA expression vector
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