Functional assessment of expression of Cre recombinase targeting the liver of transgenic mice regulated by Tet-on and Cre/loxP system
Objective To construct and evaluate transgenic mice rtTALAP-1/ LC-1 target expressing Cre recombinase in liver under regulation of Tet-on and Cre/loxP systems,to lay the foundation for construction of HCV transgenic mice model which can break the embryo stage immunotolerance under dual regulation systems.Methods Mating transgenic mice rtTALAP-1 and LC-1 to acquire offspring mice rtTALAP-1/LC-1,rtTA element and Cre gene fragment of them were detected by PCR analysis.Double positive rtTALAP-1/LC-1 mice were induced by dox for one week,then luciferase signal of mice liver were detected by small animal living body image-forming system and Cre recombinase expression in liver tissue etc.were detected by immunol histochemistry subsequently.Results Experimental results showed that clear luciferase signal can be detected only in liver of rtTALAP-1/ LC-1 mice after dox induction by small animal living body image-forming system.Immuno-histochemistry experiment results also showed that Cre recombinase expression only can be detected in liver of rtTALAP-1/ LC-1 mice,on the contrary Cre expression can not be detected in heart,kidney,skeletal muscle and other tissue of rtTALAP-1/ LC-1 mice.Conclusion Transgenic mice rtTALAP-1/ LC-1 were constructed successfully,the liver targeting and induced reaction of expression of targeted gene were very good as well.The above research has laid a solid foundation for construction of HCV transgenic mice model which can break the embryo stage immunotolerance under dual regulation systems.
Rehermann B,Nascimbeni M.Immunology of hepatitis B virus and hepatitis C virus infection[J].Nat Rev Immunol,2005,5(3):215-229. Barth H,Robinet E,Liang TJ,et al.Mouse models for the study of HCV infection and virus-host interactions[J].J Hepatology,2008,49(1):134-142. Lee JC,Tseng CK,Chen KJ,et al.A cell-based reporter assay for inhibitor screening of hepatitis C virus RNA-dependent RNA polymerase[J].Anal Biochem,2010,403(1-2):52-62. Kwong AD,McNair L,Jacobson I,et al.Recent progress in the development of selected hepatitis C virus NS3.4A protease and NS5B polymerase inhibitors[J].Curr Opin Pharmacol,2008,8(5):522-531. Bockamp E,Maringer M,Spangenberg C,et al.Of mice and models:improved animal models for biomedical research[J].Physiol Genomics,2002,11(3):115-132. Kistner A,Gossen M,Zimmermann F,et al.Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice[J].Proc Natl Acad Sci USA,1996,93(20):10933-10938. Schonig K,Schwenk F,Rajewsky K,et al.Stringent doxycycline dependent control of CRE recombinase in vivo[J].Nucleic Acids Res,2002,30(23):e134. Gossen M,Bujard H.Tight control of gene expression in mammalian cells by tetracycline-responsive promoters[J].Proc Natl Acad Sci USA,1992,89(12):5547-5551. Yu HM,Liu B,Chiu SY,et al.Development of a unique system for spatiotemporal and lineage-specific gene expression in mice[J].Proc Natl Acad Sci U S A,2005,102(24):8615-8620.
- 文章访问数: 3200
- HTML全文浏览量: 9
- PDF下载量: 1858
- 被引次数: 0