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Tools for Genomic and Transcriptomic Analysis of Microbes at Single-Cell Level

机译:单细胞水平微生物基因组和转录组学分析工具

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Microbiologists traditionally study population rather than individual cells, as it is generally assumed that the status of individual cells will be similar to that observed in the population. However, the recent studies have shown that the individual behavior of each single cell could be quite different from that of the whole population, suggesting the importance of extending traditional microbiology studies to single-cell level. With recent technological advances, such as flow cytometry, next-generation sequencing (NGS), and microspectroscopy, single-cell microbiology has greatly enhanced the understanding of individuality and heterogeneity of microbes in many biological systems. Notably, the application of multiple ‘omics’ in single-cell analysis has shed light on how individual cells perceive, respond, and adapt to the environment, how heterogeneity arises under external stress and finally determines the fate of the whole population, and how microbes survive under natural conditions. As single-cell analysis involves no axenic cultivation of target microorganism, it has also been demonstrated as a valuable tool for dissecting the microbial ‘dark matter.’ In this review, current state-of-the-art tools and methods for genomic and transcriptomic analysis of microbes at single-cell level were critically summarized, including single-cell isolation methods and experimental strategies of single-cell analysis with NGS. In addition, perspectives on the future trends of technology development in the field of single-cell analysis was also presented.
机译:传统上,微生物学家研究种群而不是单个细胞,因为通常认为单个细胞的状态将与种群中观察到的状态相似。但是,最近的研究表明,每个单个细胞的个体行为可能与整个人群的行为完全不同,这表明将传统微生物学研究扩展到单个细胞水平的重要性。随着最新技术的进步,例如流式细胞仪,下一代测序(NGS)和显微光谱学,单细胞微生物学极大地增进了人们对许多生物系统中微生物个体和异质性的理解。值得注意的是,多种“组学”在单细胞分析中的应用揭示了单个细胞如何感知,响应和适应环境,如何在外部压力下产生异质性并最终决定了整个种群的命运,以及微生物如何在自然条件下生存。由于单细胞分析不涉及目标微生物的无菌培养,因此它也已被证明是剖析微生物“暗物质”的有价值的工具。在本综述中,当前用于基因组和转录组学的最新工具和方法严格总结了单细胞水平的微生物分析,包括单细胞分离方法和NGS单细胞分析的实验策略。此外,还提出了关于单细胞分析领域技术发展的未来趋势的观点。

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