CORE--TRANSGENIC ANIMALS

该Core是一个传统的Core，并且在使用转基因小鼠进行抗镰状细胞生长策略的体内验证中具有额外的作用，这是我们最初提议的Project 1的一部分。核心服务包括三个功能：1)确保转基因和基因打靶的设施在爱因斯坦医学院的老鼠为了实现目标项目1中概述了RL内格尔将生成与DNA结构由核心的监督下B创始人埃里克Bouhassira。2)识别表达所需的血红蛋白,描述它们,并培育他们我们转基因小鼠表达α/ H /β/ S和/或新创建的老鼠RMCE目标中描述(3),alpha-knockout,β -敲除，以及其他可用的转基因和敲除系，并将其回交到C57BL/6上，用于项目1,2,4,3)为其他项目提供分析服务；蛋白质鉴定，定量（高效液相色谱，质谱）和红细胞表征。我们将利用我们现有的表征良好的模型来开发表征抗镰状红细胞凝集策略有效性的方法。开发两种新的方法来评估镰状红细胞凝集小鼠和抗镰状红细胞凝集蛋白小鼠的缺氧情况：1)HIF-1 α作为局部缺氧的检测手段。2)应用BOLD和流量敏感MRI技术监测转基因模型中脱氧血红蛋白的存在和改善/受损的血流。利用S+S+安的列斯小鼠的异常严重程度及其通过低水平γ的拯救来帮助设计更有效的抗镰状细胞生长策略。我们将利用项目2中描述的重组酶介导的盒式交换（RMCE）技术来生成一种全新的镰状细胞小鼠模型来测试抗镰状球蛋白。新的镰状细胞小鼠模型具有不同水平的转基因表达，但在小鼠的一生中拷贝数和染色质结构具有可比性，这将简化这些动物的使用。消除珠蛋白不足、地中海贫血和具有不同水平抗镰状球蛋白表达的可比小鼠将大大简化我们提出的干预措施导致的病理生理变化的解释。使用这项技术，我们期望能够明确地确定我们计划在体内环境中测试的哪种抗镰状球蛋白最好。

This Core is a traditional Core and has an additional role in in vivo validation of anti-sickling strategies using transgenic mice that formed a part of Project 1 in our original proposal. The Core services consist of three functions: 1) Make sure of the Transgenic and Gene Targeting Facility at the Albert Einstein College of Medicine where mice designed to achieve the goals outlined in Project 1 by RL Nagel will be generated with DNA constructs supplied by Core B under supervision of Eric Bouhassira. 2) Identify founders expressing the desired hemoglobins, characterize them, and breed them to our transgenic mice expression alpha/H/beta/S and/or the new mice created by RMCE (described in Aim 3), alpha-knockout, beta-knockout, and other transgenic and knockout lines available and backcross them onto C57BL/6 for use in Projects 1, 2, and 4, 3) Provide analytic services for other projects; protein identification, quantification (HPLC, mass spec), and red cell characterization. We will use our current well characterized models to develop methodologies for characterizing the efficacy of anti-sickling strategies Develop two new methods for evaluating hypoxia in sickle mice and those with anti-sickling globins: 1) HIF-1 alpha as a means of testing for local hypoxia. 2) Apply BOLD and flow sensitive MRI techniques to monitor for the presence of deoxyhemoglobin and improved/impaired flow in transgenic models. Use the anomalous severity in the S+S+Antilles mouse and its rescue by low levels of gamma to help design more effective anti-sickling strategies. We will take advantage of the Recombinase Mediated Cassette Exchange (RMCE) technology described in Project 2 to generate a completely new type of sickle cell mouse model to test anti-sickling globins. New sickle cell mouse models with variable levels of expression of the transgene but which are comparable in copy number and chromatin structure over the lifetime of the mouse will simplify use of these animals. Elimination of globin insufficiency, thalassemia, and comparable mice with various levels of anti-sickling globin expression will considerably simplify the interpretation of changes in pathophysiology resulting from our proposed interventions. Using this technology, we expect to be able to unambiguously determine which of the anti-sickling globins we are planning to test the best in an in vivo context.