人源化CD300LF抗兔瘟病毒基因修饰兔的构建

兔出血症病毒(Rabbit hemorrhagic disease virus, RHDV)引发的兔瘟，为急性、高度致死性传染病，致死率高达100%，是兔养殖业的一种毁灭性传染病。长期以来，由于该病毒不能在体外进行稳定的增殖培养，导致其分子病原学研究严重受阻。虽然大量研究已证实FUT2为该病毒的附着因子，但是申请人前期研究结果表明FUT2敲除兔不具备抗病毒能力，因此寻找新的宿主因子在抗兔瘟病毒兔的培育中尤为关键。申请人进一步的研究揭示CD300LF敲除兔可显著抑制病毒增殖，但其存在肺脏水肿的表型，无法进行封闭种群的扩繁。因此，本研究拟用人CD300LF基因的CC’和CDR3序列，替换兔的该基因与病毒结合相关的序列（CC’和CDR3），构建人源化CD300LF兔模型，以期培育抗兔瘟病毒基因修饰兔，为病毒致病机制的研究提供理想的动物模型，促进兔瘟防治药物的筛选、疫苗的研制。

Rabbit hemorrhagic disease (RHD) is caused by rabbit hemorrhagic disease virus (RHDV), which is an acute and highly fatal infectious disease with a fatality rate as high as 100%. For a long time, the study of molecular etiology of the virus has been seriously hindered, because it cannot be proliferated stably in vitro. A large number of studies have confirmed that FUT2 is the attachment factor for the virus, but applicant demonstrated that FUT2 knockout rabbits did not display antiviral ability in previous study, so it is particularly important to find new host factors for the breeding of rabbits with anti-RHDV. Previous studies have showed that CD300LF knockout rabbits could significantly inhibit the proliferation of the virus, but they generated the phenotype of pulmonary edema and could not propagate in a closed population. Therefore, we intend to replace the important sequences that bind to viruses (CC' and CDR3) of the rabbit CD300LF gene with the CC' and CDR3 sequences of human CD300LF gene to construct a humanized CD300LF rabbit model and cultivate gene modified rabbits with anti-RHDV, which could provide an ideal animal model for the study of the virus pathogenic mechanism, promote the screening of drugs for prevention and treatment of RHD,and the development of RHD vaccine.