title: Conditional gene modification in mouse liver using hydrodynamic delivery of plasmid DNA encoding Cre recombinase.
abstract: 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.
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China. HZ_Zhu@hotmail.com
authors.forename: Huan Zhang
authors.initials: HZ
authors.lastname: Zhu
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Deng Min
authors.initials: DM
authors.lastname: Feng
authors.forename: Yin
authors.initials: Y
authors.lastname: Sai
authors.forename: Jing Lun
authors.initials: JL
authors.lastname: Xue
mesh_terms: 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*
chemical_list: 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