Optimization of the base editor BE4max in chicken somatic cells.
Optimization of the base editor BE4max in chicken somatic cells.
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DOI：
10.1016/j.psj.2022.102174
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发表时间：
2022-12
期刊：
影响因子：
4.4
通讯作者：
Lu, Yangqing
中科院分区：
文献类型：
--
作者：
Xu, Tianpeng;Zhong, Jing;Huang, Zhenwen;Yu, Lintian;Zheng, Jitan;Xie, Long;Sun, Lingling;Liu, Xingting;Lu, Yangqing
先进的动物繁殖和育种生物技术使人们有可能通过直接敲入或敲除目标基因来改变性状或创造新的遗传资源。碱基编辑技术可以实现单碱基突变而不产生双链DNA断裂，是一种很有前途的用于家畜遗传修饰和育种的工具。然而，碱基编辑器（BE）在鸡中的应用尚未优化。我们评估了BE 4 max在鸡体细胞（DF-1）中的功效。对BE 4 max的关键元件胞嘧啶脱氨酶（APOBEC）进行了优化。优化后的chBE 4 max编辑器的基本编辑效率比原BE 4 max编辑器提高了10.4% ± 4.6。通过siRNA抑制鸡DF-1细胞中尿嘧啶DNA糖基化酶相关基因甲基结合结构域蛋白4（MBD 4）的表达，编辑效率比对照提高了4.43% ± 1.4。这些结果表明，该编辑器可能在家禽育种研究中有应用。
Advanced animal reproductive and breeding biotechnology has made it possible to alter traits or create new genetic resources by the direct knock-in or knock-out of target genes. Base editing technology can achieve single-base mutations without double-stranded DNA breaks, and is a promising tool for use in the genetic modification and breeding of livestock. However, the application of base editors (BEs) in chicken has not been optimized. We evaluated the efficacy of BE4max in chicken somatic cells (DF-1). The key element of BE4max, cytosine deaminase (APOBEC), was optimized for chicken. The base editing efficiency of the optimized chBE4max editor, compared with the original BE4max editor, was improved by 10.4% ± 4.6. By inhibiting the expression of the uracil DNA glycosylase-related gene methyl binding domain protein 4 (MBD4) by siRNA in chicken DF-1 cells, the editing efficiency was enhanced by 4.43% ± 1.4 compared to the control. These results suggest that this editor may have applications in poultry breeding studies.
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