Heine Heather L, Leong Hon Sing, Rossi Fabio M V, McManus Bruce M, Podor Thomas J
The James Hogg iCAPTURE Center for Cardiovascular and Pulmonary Research/MRL, University of British Columbia, St. Paul's Hospital, Vancouver, Canada.
Methods Mol Med. 2005;112:109-54. doi: 10.1007/978-1-59259-879-3_8.
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).
使用专门的报告基因来监测动物模型中实时、组织特异性的转基因表达，为规避目前与建立转基因小鼠模型相关的局限性提供了一个契机。Cre-loxP系统和四环素（Tet）诱导系统是实现条件性基因表达的有用方法，可实现空间（细胞类型特异性）和时间（诱导剂依赖性）控制。在这些诱导系统中，最常使用α-肌球蛋白重链（α-MHC）启动子，将报告基因和转基因的表达限制在心肌中。本文介绍了每个诱导系统的概况，以及用于心肌实时、非侵入性成像的推荐报告基因。诱导性基因表达系统的有效基因传递通过慢病毒载体进行，慢病毒载体具有高转导效率、长期转基因表达和低免疫原性。本章概述了将心肌特异性诱导表达系统包装到慢病毒载体中的过程，其中转基因和报告基因被转导到心肌细胞中。通过这种方式，可以利用生物发光成像（BLI）和正电子发射断层扫描（PET）监测/追踪转基因和报告基因的表达。
