Effective polyethyleneimine-mediated gene transfer into zebrafish cells.
Polyethyleneimine (PEI) has been broadly studied as a leading nonviral gene delivery carrier because of its relatively high transfection efficiency in a wide range of cell types. Here, we report gene transfer in zebrafish cells (ZF4) using PEI as a gene carrier and lipofectamine as a control. Formations of PEI-DNA complexes were characterized by a series of measurements. The particle size of PEI-DNA complexes decreased from 274 to 132 nm, the surface charge gradually increased from -26 to 29 mV, and the cytotoxicity for zebrafish cells was observed with increasing proportion of PEI. Gel retardation assay showed that DNA was completely bound by PEI with a negative-to-positive charge ratio of 4. It was observed by transmission electron microscopy that the morphology of PEI-DNA complexes was spherical with smooth surfaces. Flow cytometry revealed that the optimum transfection efficiency (27%) mediated by PEI was obtained at an negative-to-positive charge ratio of 8, which was higher than that with lipofectamine. Luciferase activity assay confirmed the increase in reporter gene expression probably due to a more efficient formation of complex between DNA and PEI than DNA and lipofectamine. In conclusion, our study demonstrates that PEI may be applied as an effective gene carrier to mediate gene transfer into zebrafish cells.
Ouyang
Sui-Dong
SD
State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen (Zhongshan) University, Guangzhou, China.
Pei
Yuan-Yuan
YY
Weng
Shao-Ping
SP
Lü
Ling
L
Yu
Xiao-Qiang
XQ
He
Jian-Guo
JG
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000235:genetics / Q000648:ultrastructure; D008055:Lipids / Q000494:pharmacology; D046529:Microscopy, Electron, Transmission; D011094:Polyethyleneimine / Q000494:pharmacology*; D014162:Transfection / Q000379:methods*; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism
D008055:Lipids; C086724:Lipofectamine; D011094:Polyethyleneimine; D004247:DNA