摘要
猪在农业和医学领域均具有重要应用价值,利用基因修饰技术能够快速提高猪的经济性状和培育人类疾病动物模型。CRISPR/Cas9基因编辑技术具有操作简单、高效和低成本的优势,在猪基因修饰领域具有重要应用。但CRISPR/Cas9易引发基因组结构不稳、大范围染色体重排和基因脱靶等问题,因而具有潜在的生物安全风险。近年基于CRISPR/Cas9开发的碱基编辑技术可以实现单个碱基定向转换,不会导致DNA双链断裂,理论上不会引发插入/缺失(Indels),对基因编辑更精准、更安全。随着碱基编辑工具的不断开发和改良,其不仅能产生C→T(A→G)突变,还能产生C、A两种碱基的同时突变以及C→G突变,增加了应用范围;改良的碱基编辑工具在保持编辑效率的同时能够降低甚至消除旁侧突变、非C→T(A→G)突变以及Indels,使得碱基编辑技术在猪遗传修饰上具有更加重要的潜在应用价值。综述了不同的碱基编辑系统原理、碱基编辑技术在基因修饰猪模型构建和猪遗传改良中的应用,以及碱基编辑系统在猪成纤维细胞中的编辑效率和脱靶情况,以期为开展猪碱基编辑应用实践提供参考。
Pigs are highly valued in both agriculture and medicine.We can quickly improve the economic properties of pigs and develop human disease models via gene modification technologies.CRISPR/Cas9 gene editing technology,with the advantages of simple operation,high efficiency and low cost,is widely applied in modifying pig genes.However,CRISPR/Cas9,due to its proneness to cause problems including structural instability of the genomes,large-scale chromosomal rearrangement and gene off-target,could pose potential risks to biological safety.In recent years,the base editing technology based on CRISPR/Cas9 can achieve the directional conversion of a single base without causing DNA double-strand breaks and insertion or loss(Indels),which is more precise and safer for gene editing.With the continuous development and improvement of base editing tools,it is not only able to produce base C→T(A→G)mutations,but also to produce simultaneous mutations of two bases(C and A)and base C→G,increasing the scope of application.improved base editing tools can reduce or even eliminate collateral mutations while maintaining editing efficiency,no C→T(A→G)mutations and Indels,making base editing technology has more important potential applications in pig genetic modification.In order to provide references for the application of base editing in pigs,the study reviews the systemic mechanism of different base editing technologies,the application of base editing technologies in the construction of genetically modified pig models and the genetic improvement of pigs as well as the editing efficiency and off-target situation of base editing systems in pig fibroblasts.
作者
盘家圣
黄秋艳
杨烨城
杨帅朋
朱向星
唐冬生
PAN Jiasheng;HUANG Qiuyan;YANG Yecheng;YANG Shuaipeng;ZHU Xiangxing;TANG Dongsheng(School of Life Science and Engineering,Foshan University/Guangdong Key Laboratory of Animal Molecular Design and Precise Breeding,Foshan 528225,China;School of Medicine,Foshan University/Guangdong Research Center of Gene Editing Engineering Technology,Foshan 528225,China)
出处
《广东农业科学》
CAS
2022年第6期118-129,共12页
Guangdong Agricultural Sciences
基金
国家重点研发计划项目子课题(2021YFA0805901-3)
国家自然科学基金(82070199)
广东省普通高校重点领域专项(2021ZDZX2050)
广东省重点领域研发计划重大科技专项(2018B020203003)。
