Knockout Of Syrian Hamster Rag1Gene Using An Efficent TLAEN And Establishment Of Monoclonel Antibody Against Syrian Hamster IFN-γ

TALEs, first found in Xanthomonas bacteria, is a natural DNA binding protein that can recognize specific DNA sequence. With this property, fusion protein containing functional domain can be used in specific binding and modification of genome DNA both in vivo and in vitro. TALEN has been widely applied to gene knock-in, knockout, transgenic anmimals, and gene therapy recently.Hamster (Golden hamster; Mesocricetus auratus), also called Syria hamsters, has served as the animal model for many human diseases. Compared with other rodent animal model, hamster has low ratio incidence of tumor, only0.5%-17%. So the hamster is a good animal model for tumor. The hamster has also been recognized as a valuable animal model for human virus diseases. For example bunyaviruses, arenavirus arboviruses, Hannibal virus and SARS. For evaluation of replication-selective oncolytic adenovirus efficacy and toxicity, the hamster is more suitable than mice. As normal tissue of mice does not support human adenovirus replication. But the hamster normal tissues such as the lungs, liver can support the replication of adenovirus.The hamster as a model animal is widely used in recent years, but it’s application in research are hampered by the limited tools, such as the transgenic hamster and related antibodies.Therefore we focus on development of genetic-modified the golden hamsters and research tools. First, we use a new gene modification tool Transcription activator-like (TAL) effectors to try to knockout the hamster Ragl gene to preparation immunodeficiency hamster. Second, Wed developed the anti-hamster IFN-y antibody.As the result, we have tested the cutting efficiency of designed TALENs targeting the hamster Ragl gene in zebrafish system. There is one pair of cotipal that can have17.9%cutting efficency in the fertilized eggs of zebrafish. We also created hybridoma which can express anti-IFN-γ mice monoclonal antibody through infusion the sp2/0with splenocyte from Balb/c mice immunized with our purified hamster IFN-γ proteins. This study provides very usefull research tools for use syrican hamster as a disease model.