Gen. of transgenic chickens using a lentiviral approach

描述(由申请人提供)： 目前，嵌合鸡是通过将囊胚层细胞或鸡胚胎干细胞注射到X期受体胚胎中而形成的，但嵌合体的频率和水平是可变的，也是不可预测的。我们正在开发一种最低限度捐献的通用接受者，这样捐赠者被剥夺的细胞对所产生的嵌合体的贡献率始终是高的和可预测的。 在该SBIR项目的第一阶段，构建了用于通用受体鸡系和GFP对照系的慢病毒载体，并用慢病毒载体转导X期鸡胚胎进行了评估。这一结果清楚地证明了慢病毒方法用于建立通用受体转基因鸡系的可行性。 在这项赠款申请的第二阶段，我们计划使用慢病毒方法产生通用受体鸡系。该项目是独特和创新的，因为它寻求建立一种可靠和可重复的方法，使受体胚胎始终产生高质量的嵌合体，并允许在短时间内生产出每个卵子大量的蛋白质。

DESCRIPTION (provided by applicant): Currently, chimeric chickens are made by injecting blastodermal cells or chicken embryonic stem cells into stage X recipient embryos, but the frequency and the level of chimerism is variable and not predictable. We are developing a universal recipient that contributes minimally, so that the contribuion of donor dervied celld to the resulting chimera is constantly high and predictable. In phase I of this SBIR project, a lentiviral vector for the universal recipient chicken line and for a GFP control line was generated and evaluated by transducing stage X chicken embryos with a lentiviral vector. The results clearly demonstrated the feasibility of a lentiviral approach for the establishment of the universal recipient transgenic chicken lines. In phase II of this grant application, we plan to generate the universal recipient chicken lines using the lentiviral approach. The project is unique and innovative because it seeks to establish a reliable and reproducible method of compromising the recipient embryos to consistently yield high-grade chimeras and allow the production of significant amounts of protein per egg in a short period of time.