REH cells and knockdowns - (Apr/24/2007 )
Has anyone knocked down anything in these dreadful cells that have given me such hell ! if so please advice on protocol as I am losing my mind with these suspension cells. thanks
The combination of Morpholino antisense delivered with Endo-Porter peptide has a good shot at working for you. We've tested Endo-Porter in suspension cells, but not in REH cells. For about a quarter of the cells tried, Endo-Porder is toxic at concentrations below that needed for delivery; however, for most cells you can get good delivery of uncharged antisense at concentrations below the onset of toxicity.
Endo-Porter is an endosomal release agent. After co-endocytosis of uncomplexed neutral-charge antisense and Endo-Porter peptide, acidification of the endosome protonates Endo-Porter and in its cationic form it makes the endosomal membrane leaky, allowing large molecules to enter the cytosol.
Good luck with your uncooperative cells!