SCID兔和类NOD-SCID兔的制备与应用

目前，原发性免疫缺陷病在发病机制和治疗措施的研究上依然面临困难。虽然，免疫缺陷小鼠模型可以模拟人免疫缺陷疾病的部分表型，作为动物模型也广泛用于免疫疾病、肿瘤、传染病以及造血机制等生物医学研究中。但是，小鼠与人种属差异较大，免疫缺陷小鼠并不能完全反映人免疫系统缺陷导致的相关疾病发病过程。由于兔在免疫系统基因表达上与人类更接近，免疫缺陷兔模型将更好地模拟人免疫缺陷疾病发生、发展过程。本项目将分别利用CRISPR/Cas9技术敲除兔Prkdc基因以及在兔Rosa26位点插入人SPIRa基因，获得两种基因修饰兔，经过繁殖获得SCID和类NOD-SICD两种免疫缺陷兔。通过分析SCID、类NOD-SICD兔的免疫缺陷表型，确定免疫缺陷的类型；并利用此模型进行细胞异体移植和免疫系统人源化兔的构建。此基因修饰兔为临床上深入研究免疫缺陷病及相关疾病发病机制、开发治疗方案、筛选靶向药物提供实用的动物模型。

The present research on the pathogenesis and effective treatment of primary immunodeficiency is still difficult. The immunodeficient mice can simulate part of the phenotype of human immunodeficiency disease and be wildly used in biomedical research of immune disease, tumor, infectious disease and hematopoietic mechanism as well. However, due to the large differences between mice and human species, the immunodeficient mice cannot fully reflect the pathogenesis of diseases caused by human immune system defects. As the expression of immune system genes in rabbits is closer to that in humans, immunodeficient rabbit model will better mimic the general pattern of the occurrence and development of human immunodeficiency diseases. This project will use the CRISPR/Cas9 technology to knock out rabbit Prkdc gene and insert the human SPIRa gene at rabbit Rosa26 site to obtain two genetically modified rabbits, and obtain SCID rabbits and NOD-SCID like immunodeficient rabbits through natural breeding. Immunodeficiency phenotype of two kinds of immunodeficient rabbits will be analyzed to confirm the types of immunodeficiency. In addition, the models will be used for allografts and the construction of humanized rabbits. The genetically modified rabbits will provide a practical animal model for in-depth study of the pathogenesis of immunodeficiency and other related disease, development of therapeutic measures and screening of targeted drugs.