Development of single-cell sequencing technology for microbial populations

微生物群体单细胞测序技术的发展

• 批准号: BB/R022526/1
• 负责人: Neil Hall
• 金额: $ 19.06万
• 依托单位: Earlham Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR022526%2F1
• 关键词: Development; single; cell; sequencing; technology

When biological samples are analysed at the DNA level (to study genotypes) or the RNA level) to look at gene expression. We currently have to analyze many thousands or millions of cells at a time. This has been a very informative approach but it means that we miss a lot of complexity in biological systems, as in most cases the DNA in a microbial culture may be different in each cell and the gene expression in each cell is also hetrogeneous. This is particularly important is communities that form biofilms on surfaces as there may be different strains or species of bacteria working together performing different roles in the community. Biofilms are especially important for bacteria evading antimicrobial treatment. In this project we aim to develop the methodologies for single cell sequencing of microbes and test these against a number of different use cases. We will use Fluorescence Activated Cell Sorting (FACS) to separate cells and then develop protocols to analyze these cells in parallel by using molecular barcoding methods.

当生物样本在DNA水平（研究基因型）或RNA水平上进行分析以观察基因表达时。目前，我们必须一次分析数千或数百万个细胞。这是一种非常有用的方法，但这意味着我们错过了生物系统的许多复杂性，因为在大多数情况下，微生物培养中的DNA在每个细胞中可能是不同的，每个细胞中的基因表达也是异质的。这对于在表面形成生物膜的群落尤其重要，因为可能有不同的菌株或种类的细菌在群落中共同发挥不同的作用。生物膜对逃避抗菌处理的细菌尤其重要。在这个项目中，我们的目标是开发微生物单细胞测序的方法，并针对许多不同的用例对这些方法进行测试。我们将使用荧光活化细胞分选（FACS）分离细胞，然后制定方案，分析这些细胞并行使用分子条形码方法。

项目成果

Bacterial single-cell genomics enables phylogenetic analysis and reveals population structures from in vitro evolutionary studies

细菌单细胞基因组学能够进行系统发育分析，并通过体外进化研究揭示种群结构

• DOI: 10.1101/2020.08.25.266213
• 发表时间: 2020
• 作者: Bawn M

Single-cell genomics reveals population structures from in vitro evolutionary studies of Salmonella.

• DOI: 10.1099/mgen.0.000871
• 发表时间: 2022-09
• 期刊: MICROBIAL GENOMICS
• 影响因子: 3.9
• 作者: Bawn, Matt;Hernandez, Johana;Trampari, Eleftheria;Thilliez, Gaetan;Quince, Christopher;Webber, Mark A.;Kingsley, Robert A.;Hall, Neil;Macaulay, Iain C.
• 通讯作者: Macaulay, Iain C.