Development of a Recombineering System for the Acetogen <i>Eubacterium limosum</i> with Cas9 Counterselection for Markerless Genome Engineering.

ABSTRACT: Eubacterium limosum is a Clostridial acetogen that efficiently utilizes a wide range of single-carbon substrates and contributes to metabolism of health-associated compounds in the human gut microbiota. These traits have led to interest in developing it as a platform for sustainable CO₂-based biofuel production to combat carbon emissions, and for exploring the importance of the microbiota in human health. However, synthetic biology and metabolic engineering in E. limosum have been hindered by the inability to rapidly make precise genomic modifications. Here, we screened a diverse library of recombinase proteins to develop a highly efficient oligonucleotide-based recombineering system based on the viral recombinase RecT. Following optimization, the system is capable

SUBMITTER: Sanford PA 

PROVIDER: S-EPMC11334238 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature