High efficient AAV-transducible transgenic quails

项目摘要/摘要 这项研究计划的主要目标是产生一个普遍表达这种基因的转基因鹌鹑品系。 人腺相关病毒受体(AAV)。不能用ffi对鹌鹑和鸡进行基因转换 AAV，这排除了在这些动物中使用这种流行病毒的可能性。AAV受体的表达 然而，在禽类细胞中，fi显著增加了对甲型肝炎病毒的转导ffi的敏感性。建议的fi的首要目标 研究是为了产生一种转基因的鹌鹑品系，可以测试其体内是否存在ffi病毒的转导 耳在活体内，这是目标二的提议。fiNAL的目标是证明新的 构建的表达AAV受体的转基因鹌鹑品系可用于AAV介导的CRISPR-Cas9 在内耳进行基因组编辑。这项研究项目的成功完成将提供一种新的动物 通过体内基因操作研究禽类毛细胞再生的模型。内耳感觉毛细胞 哺乳动物不会自然再生，但鸟类会，这就强调了传导的重要性 鸟类的基因操作。通过这个新的实验模型系统获得的知识，例如 在鸟类毛细胞再生过程中，特殊的fic信号通路和关键基因的信号转导应该是 可翻译给哺乳动物，最终可以为感音神经性听力的治疗提供洞察力 损失。

Project Summary / Abstract The main goal of this research proposal is to generate a transgenic quail line that ubiquitously expresses the human receptor for adeno-associated virus (AAV). Quails and chickens cannot be efficiently transduced with AAV, which precludes the use of this class of popular viruses in these animals. Expressing the AAV receptor in avian cells, however, significantly increases the transduction efficacy for AAV. The first aim of the proposed research is to generate a transgenic quail line that can be tested for efficient virus transduction of the inner ear in vivo, which is the goal of aim two of the proposal. The final aim is to demonstrate that the newly generated transgenic AAV receptor-expressing quail line can be used for AAV-mediated CRISPR-Cas9 genome editing in the inner ear. The successful completion of this research project will provide a new animal model for studying avian hair cell regeneration through gene manipulation in vivo. Inner ear sensory hair cells do not naturally regenerate in mammals but they do in birds, which emphasizes the importance of conducting genetic manipulations in birds. Knowledge gained with this new experimental model system, such as the significance of specific signaling pathways and key genes during avian hair cell regeneration, shall be translatable to mammals, and ultimately can provide insight into curative treatments for sensorineural hearing loss.