摘要
目的制作载脂蛋白E（ApoE）基因敲除（ApoE^（-/-））兔,为研究动脉粥样硬化提供优良动物模型。方法应用CRISPR/Cas9系统编辑新西兰兔的Apo E基因,制作F0代ApoE^（-/-）兔,利用PCR、TA克隆、Sanger测序、Western blot和血脂检测鉴定Apo E靶向敲除的效率和效果,并通过传代获得ApoE^（＋/-）兔。结果 Sanger测序结果显示成功靶向敲除兔ApoE基因,血清脂蛋白含量在正常饮食状态下相对于野生型兔有升高,且可稳定遗传。结论成功建立了ApoE^（-/-）兔,为进一步研究动脉粥样硬化和其他相关疾病提供了良好的动物模型。
Aim To provide an excellent animal model for the study of atherosclerosis,it generated apolipoprotein E gene knockout（ ApoE^（-/-）） rabbit. Methods Apo E gene of New Zealand rabbits were edited and F0 ApoE^（-/-）rabbits were created by applying CRISPR/Cas9 system. The efficiency and effect of Apo E gene targeted knockout was determined through PCR,TA cloning,Sanger sequencing,Western blot and lipid assay. Apo E＋/-rabbits were obtained through transmitting by germline. Results Sanger sequencing results showed that Apo E gene was successfully targeted knockout. Serum lipoprotein content of ApoE^（-/-）rabbits increased in normal diet compared with wild type rabbits and can be transmitted by germline. Conclusion The successful establishment of ApoE^（-/-）rabbit provided a good animal model for the further study of atherosclerosis and other related diseases.
作者
袁婷婷
周敏雅
卢瑶瑶
陆睿
张婷
孙雨婷
钟港
王梦恬
黄俊杰
胡艳华
梁景岩
成勇
YUAN Tingting;ZHOU Minya;LU Yaoyao;LU Rui;ZHANG Ting;SUN Yuting;ZHONG Gang;WANG Meng-tian;HUANG Junjie;HU Yanhua;LIANG Jingyan;CHENG Yong(Jiangsu Key Laboratory of Senile Diseases Control and Prevention with Integrated Traditional Chinese and Western Medi-cine,Medical College of Yangzhou University,Yangzhou,Jiangsu 225001;Jiangsu Provincial Research Center for Animal Transgencsis and Biopharming,College of Veterinary Medicine Yangzhou University,Yangzhou,Jiangsu 225009,China)
出处
《中国动脉硬化杂志》
CAS
2018年第7期658-665,共8页
Chinese Journal of Arteriosclerosis
基金
国家重点研发计划(2016YFE0126000)
国家自然科学基金(81570392)
扬州大学研究生科研创新计划项目(XKYCX17_069)
