基于CRIPSR/Cas9全基因组扫描文库筛选羊布氏杆菌宿主互作基因及功能验证

布氏杆菌病是一种由布氏杆菌引起的动物源性人畜共患病，不仅给整个畜牧业造成了巨大的经济损失，也同样严重威胁着人类的健康。目前布氏杆菌的致病机制尚不明确，导致疫苗和治疗药物的开发以及抗病育种研究进展缓慢。针对此科学问题，本项目首先利用近年来兴起的CRISPR/Cas9全基因组扫描技术，在人源易感细胞系中从全基因组水平高通量筛选与布氏杆菌侵染和复制相关的宿主因子，并在羊源免疫细胞中对筛选到的关键宿主基因进一步验证；其次，构建布氏杆菌/宿主因子互作网络，阐明关键宿主因子调控布氏杆菌在宿主细胞（人和羊）内复制的分子机制；最后，基于本课题组已建立的基因编辑羊高效制备体系，创制抗布氏杆菌感染的基因编辑羊模型，并在活体水平上开展抗病能力评价。该研究将为抗布氏杆菌药物的开发和抗布病育种提供重要的靶点，也将为明晰布氏杆菌的致病机理提供重要线索，对保障人类健康及健康养殖的跨越式发展具有重要意义。

Brucellosis is a zoonosis caused by Brucella, which not only causes huge economic losses to the entire animal husbandry, but also seriously threatens human health. At present, the pathogenic mechanism of Brucella is still unclear, which cause the slow progress in the development of vaccines and therapeutic medicines, and research on disease resistance breeding. To address this scientific issue, we firstly use a CRISPR/Cas9 genome-wide sgRNA library to screening of the host factors related to Brucella infection and replication in human susceptible cell lines and verifying the candidate host gene in sheep-derived immune cells. Secondly, constructing a Brucella/host factors interaction network and illuminating the molecular mechanism of several key host factors regulating the replication of Brucella in host cells (human and sheep). Finally, basing on the efficient generation of genome-edited sheep system in our group, creating the anti-brucella infection genome-edited sheep model, and carrying out disease resistance evaluation. This research will provide a crucial targets for the development of anti-brucellosis drugs and disease resistance breeding. Meanwhile, the research may offer important clues that clarify the pathogenic mechanism of brucella, which is significant for protecting human health and the leap-forward development of healthy breeding.