Liu Ya-Jun, Zhang Jie, Cui Gu-Zhen, Cui Qiu
Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, P.R. China.
Qingdao Engineering Laboratory of Single Cell Oil, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, P.R. China.
Biotechnol J. 2015 Jun;10(6):855-65. doi: 10.1002/biot.201400716. Epub 2015 Mar 3.
Targetrons are mobile group II introns that can recognize their DNA target sites by base-pairing RNA-DNA interactions with the aid of site-specific binding reverse transcriptases. Targetron technology stands out from recently developed gene targeting methods because of the flexibility, feasibility, and efficiency, and is particularly suitable for the genetic engineering of difficult microorganisms, including cellulolytic bacteria that are considered promising candidates for biomass conversion via consolidated bioprocessing. Along with the development of the thermotargetron method for thermophiles, targetron technology becomes increasingly important for the metabolic engineering of industrial microorganisms aiming at biofuel/chemical production. To summarize the current progress of targetron technology and provide new insights on the use of the technology, this paper reviews the retrohoming mechanisms of both mesophilic and thermophilic targetron methods based on various group II introns, investigates the improvement of targetron tools for high target efficiency and specificity, and discusses the current applications in the metabolic engineering for bacterial producers. Although there are still intellectual property and technical restrictions in targetron applications, we propose that targetron technology will contribute to both biochemistry research and the metabolic engineering for industrial productions.
归巢核酸内切酶是可移动的II类内含子,借助位点特异性结合逆转录酶,通过RNA-DNA碱基配对相互作用识别其DNA靶位点。归巢核酸内切酶技术因灵活性、可行性和效率,在近期开发的基因靶向方法中脱颖而出,特别适用于对难处理微生物进行基因工程改造,包括被视为通过整合生物加工进行生物质转化的有前景候选菌的纤维素分解菌。随着嗜热菌的嗜热归巢核酸内切酶方法的发展,归巢核酸内切酶技术对于旨在生产生物燃料/化学品的工业微生物代谢工程变得越来越重要。为总结归巢核酸内切酶技术的当前进展并为该技术的应用提供新见解,本文综述了基于各种II类内含子的嗜温和嗜热归巢核酸内切酶方法的逆转归巢机制,研究了提高归巢核酸内切酶工具靶向效率和特异性的方法,并讨论了其在细菌生产者代谢工程中的当前应用。尽管归巢核酸内切酶应用仍存在知识产权和技术限制,但我们认为归巢核酸内切酶技术将有助于生物化学研究和工业生产的代谢工程。