Xie Tian, Wang Mei, Gao Rui-Yu, Miao Yan-Ni, Zhang Yi-Ming, Jiang Jing
Genome Tagging Project Center, Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
Yi Chuan. 2022 Aug 20;44(8):655-671. doi: 10.16288/j.yczz.22-158.
The site-specific recombination systems are composed of recombinases and specific recognition sites, which are powerful tools for gene manipulation and have been extensively used in life sciences research. Inducible recombination systems have been developed to precisely regulate gene expression in a spatiotemporal manner in cells and animals for applications such as gene function research, cell lineage tracing and disease treatment. Based on different spatiotemporal expression methods of recombinases, inducible recombination systems can be divided into two categories: chemical- controlled and light-controlled inductions. Light-controlled inducible recombination systems that utilize light as inducer consist of photocage and optogenetics in accordance with optical control patterns and objects. Photocaged inducible recombination systems are using photosensitive groups to control chemical inducers or recombinases. Their activities are inhibited by photosensitive groups before light induction and recovered after specific light irradiation, leading to light-controlled inducible gene recombination. While optogenetic inducible recombination systems rely on reactivations of split recombinases that mediated by optogenetic switches. Optogenetic switches are composed of a series of gene-encoded photosensitive proteins, including cryptochromes, VIVID, phytochromes, etc. These types of light-controlled inducible recombination systems provide more possibilities for analyzing gene expression and function from the dimension of high spatiotemporal resolution to meet the increasingly complex demands of life science research. In this review, we summarize the developing principles and applications of different types of light-controlled inducible recombination systems, compare their advantages and disadvantages, and prospect the development of more light-controlled recombination systems in the future, with the aims to provide theoretical basis and guidance for system optimization and upgrade.
位点特异性重组系统由重组酶和特定识别位点组成，是基因操作的强大工具，已在生命科学研究中广泛应用。诱导型重组系统已被开发出来，用于在细胞和动物中以时空方式精确调控基因表达，应用于基因功能研究、细胞谱系追踪和疾病治疗等领域。根据重组酶不同的时空表达方式，诱导型重组系统可分为两类：化学诱导和光诱导。利用光作为诱导剂的光控诱导型重组系统根据光学控制模式和对象可分为光笼和光遗传学。光笼诱导型重组系统利用光敏基团来控制化学诱导剂或重组酶。在光诱导之前，它们的活性被光敏基团抑制，在特定光照射后恢复，从而实现光控诱导型基因重组。而光遗传学诱导型重组系统则依赖于由光遗传学开关介导的分裂重组酶的重新激活。光遗传学开关由一系列基因编码的光敏蛋白组成，包括隐花色素、VIVID、光敏色素等。这些类型的光控诱导型重组系统从高时空分辨率维度为分析基因表达和功能提供了更多可能性，以满足生命科学研究日益复杂的需求。在本综述中，我们总结了不同类型光控诱导型重组系统的发展原理和应用，比较了它们的优缺点，并展望了未来更多光控重组系统的发展，旨在为系统优化和升级提供理论依据和指导。
