Increasing the targeting scope and efficiency of base editing with Proxy-BE strategy
Increasing the targeting scope and efficiency of base editing with Proxy-BE strategy
复制标题
使用 Proxy-BE 策略提高碱基编辑的靶向范围和效率
DOI：
10.1002/1873-3468.13719
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发表时间：
2019
期刊：
影响因子：
3.5
通讯作者：
Wei Yongchang
中科院分区：
文献类型：
--
作者：
Liu Yin;Li Guanglei;Yang Guang;Gu Huifeng;Huang Shisheng;Yu Wenxia;Qin Guizhen;Liu Xinyi;Zhou Fuling;Huang Xingxu;Wei Yongchang
碱基编辑器因其简单、通用性强而广泛应用于精确基因编辑。然而，它们的效率受到各种障碍的阻碍。考虑到染色质微环境是一个可能的障碍，在这里，我们进一步开发了proxy - clustered regularly interspaced short palindromic repeats策略，称为proxy - BE，以提高基因编辑效率。具体来说，一个核酸酶死亡的Cas9 （dCas9）被结合到距离目标位点约20-30个碱基对的序列上，这可能会改善对DNA的访问，从而为碱基编辑器提供更好的编辑微环境。我们的研究结果证实，核酸酶-死亡化脓链球菌cas9可以帮助碱基编辑器SaKKH - BE3和dCpf1 - BE将其标准碱基编辑效率提高一倍。这项工作为加强碱基编辑提供了新的途径，扩大了其在生物学研究和基因治疗中的应用范围。
Base editors (BEs) are widely used in precise gene editing due to their simplicity and versatility. However, their efficiencies are hindered by various obstacles. Considering the chromatin microenvironment as a possible obstacle, here, we demonstrate a further development of the proxy‐clustered regularly interspaced short palindromic repeats strategy, termed Proxy‐BE, to increase gene editing efficiency. Specifically, a nuclease‐dead Cas9 (dCas9) was bound to the sequence about 20–30 base pair away from the target site, potentially improving access to the DNA and, thus, providing a better editing microenvironment for base editors. Our findings confirm that nuclease‐deadStreptococcus pyogenesCas9 can assist the base editors SaKKH‐BE3 and dCpf1‐BE to double their canonical base editing efficiency. This work provides a new approach to enhance base editing, extending its scope for biological research and gene therapy.