In vitro and in vivo functional assays

Core B提供体外和体内测定骨密度样肌营养不良蛋白的生化功能。这些内部缺失的半功能蛋白是由其他研究项目和核心产生的，包括对具有良好特征的框架内突变的患者的人类细胞中的内源性蛋白的研究（项目1,3），以vh/o传递给细胞或小鼠的特定缺失的小鼠构建（项目1），以及通过外显子跳跃疗法完成的框架内缺失（项目2）。核心B的具体目标包括由国家儿童医学中心遗传医学研究中心内的五个合作实验室开发的完善的分析。Core B提供的一项创新检测是我们最近发表的一种活体成像方法，用于使用近红外组织蛋白酶B笼底物评估肌肉萎缩症活动（Baudy et al 2010）。另一项创新分析是使用MS/MS分析定量分泌组测量膜不稳定性。

Core B provides in vitro and in vivo assays for determining the biochemical functionality of BMD-like dystrophin proteins. These internally deleted semi-functional proteins are generated by the other research projects and cores, and include studies of endogenous proteins in human cells from patients with wellcharacterized in-frame mutations (Projects 1, 3), murine constructs of specific deletions delivered to cells or mice in vh/o (Project 1), and in-frame deletions accomplished by exon-skipping therapeutics (Project 2). The specific aims of Core B include well-established assays developed by the five collaborating laboratories within the Research Center for Genetic Medicine at Children's National Medical Center. An innovative assay offered by Core B is our recently published live-animal imaging method for assessment of muscular dystrophy activity using a near-infrared cathepsin B caged substrate (Baudy et al 2010). An additional innovative assay is quantitative secretome measures of membrane instability using MS/MS profiling. Resource sharing will include publication and public access of standard operating procedures for each assay, as we have done collaboratively with the EU Treat-NMD network for murine pre-clinical endpoints (http://www.treat-nmd.eu/research/preclinical/SOPs/) (Nagaraju 2009; Spurney et al. 2009). We also offer in vivo assays as a core function for drug screening to external laboratories, and in vitro assays to external laboratories on a collaborative basis.