产甲烷古细菌,它们是全球碳和氮循环的组成部分,由于独特的生理学和有限的遗传工具,目前在遗传操作方面面临挑战。这篇综述概述了产甲烷菌基因工程的当前和过去的发展,包括选择和反选择标记,报告系统,穿梭矢量,诱变方法,无标记遗传交换,和基因表达控制。这篇综述讨论了遗传工具,并强调了与特定产甲烷物种的工具稀缺相关的挑战。产甲烷菌的诱变技术,包括物理化学,转座子介导的,脂质体介导的诱变,和自然的转变,概述,成就和挑战。无标记基因交换策略,如同源重组和CRISPR/Cas介导的基因组编辑,也是详细的。最后,该综述通过研究产甲烷菌基因表达的控制来得出结论。所提供的信息强调了在考古研究中迫切需要完善的遗传工具。尽管面临历史挑战,最近的进步,特别是基于CRISPR的系统,承诺克服障碍,对全球健康的影响,农业,气候变化,和环境工程。这篇全面的综述旨在弥合文献中现有的差距,指导未来考古基因工程领域的研究。
Methanogenic archaea, which are integral to global carbon and nitrogen cycling, currently face challenges in genetic manipulation due to unique physiology and limited genetic tools. This review provides a survey of current and past developments in the genetic engineering of methanogens, including selection and counterselection markers, reporter systems, shuttle vectors, mutagenesis methods, markerless genetic exchange, and gene expression control. This review discusses genetic tools and emphasizes challenges tied to tool scarcity for specific methanogenic species. Mutagenesis techniques for methanogens, including physicochemical, transposon-mediated, liposome-mediated mutagenesis, and natural transformation, are outlined, along with achievements and challenges. Markerless genetic exchange strategies, such as homologous recombination and CRISPR/Cas-mediated genome editing, are also detailed. Finally, the review concludes by examining the control of gene expression in methanogens. The information presented underscores the urgent need for refined genetic tools in archaeal research. Despite historical challenges, recent advancements, notably CRISPR-based systems, hold promise for overcoming obstacles, with implications for global health, agriculture, climate change, and environmental engineering. This comprehensive review aims to bridge existing gaps in the literature, guiding future research in the expanding field of archaeal genetic engineering.