嗜热厌氧杆菌热稳定CRISPR/Cas9基因组编辑技术及在细胞工厂构建中的应用研究进展

Thermostable CRISPR/Cas9 genome editing system and its application in construction of cell factories with thermophilic bacteria:a review

嗜热厌氧杆菌属(Thermoanaerobacter)来源的菌株作为一个高温发酵的细胞工厂,具有生产生物燃料和化学品的潜力,已引起了研究者的广泛兴趣。随着嗜热厌氧杆菌属来源的多个菌株全基因组测序的完成以及相关的生理生化实验的开展,建立一个简便、快捷的基因操作技术将有助于进一步深入理解和认识嗜热厌氧杆菌有关的代谢途径。最近,成簇的规律间隔短回文重复序列及其相关系统(CRISPR/Cas)的基因编辑技术在嗜热菌中被开发应用。文中综述了嗜热厌氧杆菌的研究进展和热稳定性CRISPR/Cas9基因组编辑技术在嗜热厌氧杆菌的发展和建立,并对该编辑技术在其他嗜热菌未来发展的趋势提出参考意见。嗜热菌热稳定性的CRISPR/Cas9基因组编辑技术的建立和完善不仅可以促进嗜热菌遗传改造技术的发展,而且有助于提高嗜热菌遗传育种和代谢工程改造的效率。

The diverse thermophilic strains of Thermoanaerobacter,serving as unique platforms with a broad range of application in biofuels and chemicals,have received wide attention from scholars and practitioners.Although biochemical experiments and genome sequences have been reported for a variety of Thermoanaerobacter strains,an efficient genetic manipulation system remains to be established for revealing the biosynthetic pathways of Thermoanaerobacter.In line with this demand,the clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated(Cas)systems for editing,regulating and targeting genomes have been well developed in thermophiles.Here,we reviewed and discussed the current status,associated challenges,and future perspectives of the construction of thermostable CRISPR/Cas9 genome editing systems for some representative Thermoanaerobacter species.The establishment,optimization,and application of thermostable CRISPR/Cas genome editing systems would potentially provide a foundation for further genetic modification of thermophilic bacteria.