Characterisation and enhancement of cyanobacterial extracellular electron transfer using CRISPR tools and photoelectrochemical analysis

蓝藻将电子传输到细胞外环境的能力(称为外生电活性)已被用来构建能够太阳能发电的生物光伏装置。然而，对这一活动背后的生物过程的有限了解阻碍了对高度外生电致蓝藻菌株的基因工程，这将提高这些设备产生的功率密度。该项目旨在识别其产品增强或抑制外生电活性的基因，并利用这些信息来设计一种高度外电产生的蓝藻菌株。SgRNA文库将用于利用CRISPRi建立突变蓝藻的单基因敲除文库。文库成员的外生电活性将使用定制的电极微阵列进行筛选。在此之后，将开发一个通用的CRISPR-Cas9载体来促进蓝藻的基因组编辑，以产生高度外生电子的菌株。

The ability of cyanobacteria to transport electrons to their extracellular environment (termed exoelectrogenic activity) has been exploited to construct biophotovoltaic devices capable Of solar power generation. However, a limited understanding of the biological processes underlying this activity prevent genetic engineering of highly exoelectrogenic strains of cyanobacteria which would boost the power densities produced by these devices. This project aims to identify genes whose products enhance or inhibit exoelectrogenic activity and use this information to engineer a highly exoelectrogenic cyanobacterial strain. sgRNA libraries will be used to generate single-gene knockdown libraries of mutant cyanobacteria using CRISPRi. The exoelectrogenic activity of library members will be screened using a custom electrode microarray. Following this, a general-purpose CRISPR-Cas9 vector will be developed to facilitate genome editing of cyanobacteria to produce a highly exoelectrogenic strain.