2017-2020年吉林长春基于CRISPR/Cas9技术高效构建犬瘟热重组金丝雀痘病毒疫苗及大熊猫免疫研究数据集

将Purevax? Ferret Distemper进行一定的改造，使其同时表达出犬瘟热病毒的M、F和H蛋白，并组装出CDV-VLPs，以期能够提高疫苗的免疫原性，为免疫其他物种动物提供新的疫苗候选株。而由于传统的同源重组法构建重组病毒成功率低，子代病毒中重组病毒所占比例极小，筛选难度高，耗时长，因此探索出更高效的重组利用CRISPR/Cas9技术成功开发出一套快速构建重组金丝雀痘病毒的程序，并以此方法构建了同时表达犬瘟热病毒的M、F和H蛋白，并组装形成CDV-VLPs的重组病毒，小鼠、犬、狐狸和水貂免疫证明该重组病毒的免疫效果良好，探索了重组病毒最佳的免疫程序，并尝试进行了大熊猫的本动物免疫研究，为候选疫苗的选择提供了数据支持。

We modified the Purevax® Ferret Distemper vector to simultaneously express the M, F, and H proteins of canine distemper virus (CDV), and assembled CDV virus-like particles (CDV-VLPs), aiming to enhance the vaccine's immunogenicity and provide a novel vaccine candidate strain for immunizing other animal species. Traditional homologous recombination methods for constructing recombinant viruses have low success rates, with only a tiny proportion of recombinant viruses present in the progeny, accompanied by high screening difficulty and long experimental duration. To overcome these drawbacks, we developed a highly efficient and rapid protocol for constructing recombinant canarypox viruses using CRISPR/Cas9 technology. Using this method, we constructed a recombinant canarypox virus that simultaneously expresses the M, F, and H proteins of CDV and assembles into CDV-VLPs. Immunization experiments in mice, dogs, foxes, and minks demonstrated that this recombinant virus exhibits excellent immune efficacy. We also explored the optimal immunization program for this recombinant virus, and conducted preliminary immunization studies on giant pandas as their natural host, providing data support for the selection of the candidate vaccine.