Conditional gene modification in mouse liver using hydrodynamic delivery of plasmid DNA encoding Cre recombinase.
The success of Cre-mediated conditional gene targeting in liver of mice has until now depended on the generation of Cre recombinase transgenic mice or on viral-mediated transduction. Here, we sought to establish the feasibility of using hydrodynamic gene delivery of Cre recombinase into liver, using a ROSA26 EGFP mouse. The expression of EGFP and beta-galactosidase was exclusively detected in the liver of mice treated with hydrodynamic gene delivery of Cre recombinase, as assessed with fluorescence microscopy and X-Gal staining, respectively; Southern blotting also showed that Cre mediated recombination occurred specifically in the liver and not in other organs. The Cre mediated recombination reached about 61% of hepatocytes of mouse after repeated injection, as analyzed by flow cytometry. These results demonstrate that Cre recombinase can be transferred to the liver of mice through a simple hydrodynamic gene-delivery approach and can mediate efficient recombination in hepatocytes. Thus, hydrodynamic gene delivery of the Cre recombinase provides a valuable approach for Cre-loxP-mediated conditional gene modification in the liver of mice.
Zhu
Huan Zhang
HZ
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China. HZ_Zhu@hotmail.com
Wang
Wei
W
Feng
Deng Min
DM
Sai
Yin
Y
Xue
Jing Lun
JL
D000818:Animals; D004247:DNA / Q000008:administration & dosage; D005456:Fluorescent Dyes / Q000032:analysis; D018390:Gene Targeting; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage; D011506:Proteins / Q000235:genetics; D022661:RNA, Untranslated; D011995:Recombination, Genetic; D014162:Transfection / Q000379:methods*
D005456:Fluorescent Dyes; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D011506:Proteins; D022661:RNA, Untranslated; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA; C045073:Cre recombinase; D019426:Integrases