On the disulfide-linker strategy for designing efficacious cationic transfection lipids: an unexpected transfection profile.
Herein, employing a previously reported disulfide-linker strategy, we have designed and synthesized a novel cationic lipid 2 with a disulfide-linker and its non-disulfide control analog lipid 1. The relative efficacies of lipids 1 and 2 in transfecting CHO, COS-1 and MCF-7 cells were measured using both reporter gene and whole cell histochemical staining assays. In stark contrast to the expectation based on the disulfide-linker strategy, the control non-disulfide cationic lipid 1 showed phenomenally superior in vitro transfection efficacies to its essentially transfection incompetent disulfide counterpart lipid 2. Results in DNase I protection experiments and the electrophoretic gel patterns in the presence of glutathione, taken together, are consistent with the notion that the success of the disulfide-linker strategy may depend more critically on the DNase I sensitivity of the lipoplexes than on the efficient DNA release induced by intracellular glutathione pool.
Division of Lipid Science and Technology, Indian Institute of Chemical Technology, Hyderabad 500 007, India.
Valluripalli Vinod
VV
Kumar
Arabinda
A
Chaudhuri
D000818:Animals; D001943:Breast Neoplasms; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003850:Deoxyribonuclease I / Q000378:metabolism; D004220:Disulfides / Q000138:chemical synthesis*; D015195:Drug Design; D006801:Humans; D007202:Indicators and Reagents; D014162:Transfection / Q000379:methods*
D004220:Disulfides; D007202:Indicators and Reagents; D003850:Deoxyribonuclease I