Strategies of conditional gene expression in myocardium: an overview.
The use of specialized reporter genes to monitor real-time, tissue-specific transgene expression in animal models offers an opportunity to circumvent current limitations associated with the establishment of transgenic mouse models. The Cre-loxP and the tetracycline (Tet)-inducible systems are useful methods of conditional gene expression that allow spatial (cell-type-specific) and temporal (inducer-dependent) control. Most often, the alpha-myosin heavy chain (alpha-MHC) promoter is used in these inducible systems to restrict expression of reporter genes and transgenes to the myocardium. An overview of each inducible system is described, along with suggested reporter genes for real-time, noninvasive imaging in the myocardium. Effective gene delivery of the inducible gene expression system is carried out by lentiviral vectors, which offer high transduction efficiency, long-term transgene expression, and low immunogenicity. This chapter outlines the packaging of myocardium-specific inducible expression systems into lentiviral vectors, in which a transgene and a reporter gene are transduced into cardiomyocytes. In doing so, transgene and reporter expression can be monitored/tracked with bioluminescence imaging (BLI) and positron emission tomography (PET).
Heine
Heather L
HL
The James Hogg iCAPTURE Center for Cardiovascular and Pulmonary Research/MRL, University of British Columbia, St. Paul's Hospital, Vancouver, Canada.
Leong
Hon Sing
HS
Rossi
Fabio M V
FM
McManus
Bruce M
BM
Podor
Thomas J
TJ
D000818:Animals; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D019426:Integrases / Q000235:genetics; D007763:Lac Operon; D016086:Lentivirus / Q000235:genetics; D008156:Luciferases / Q000378:metabolism; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D049268:Positron-Emission Tomography; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D019076:Transgenes; D014764:Viral Proteins / Q000235:genetics
D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases; D013937:Thymidine Kinase; C045073:Cre recombinase; D019426:Integrases; D013752:Tetracycline