Genome streamlining to improve performance of a fast-growing cyanobacterium Synechococcus elongatus UTEX 2973

Annesha Sengupta, Anindita Bandyopadhyay, Debolina Sarkar, John I. Hendry, Max G. Schubert, Deng Liu, George M. Church, Costas D. Maranas, Himadri B. Pakrasi

Research output: Contribution to journalArticlepeer-review

Abstract

Cyanobacteria are photosynthetic organisms that have garnered significant recognition as potential hosts for sustainable bioproduction. However, their complex regulatory networks pose significant challenges to major metabolic engineering efforts, thereby limiting their feasibility as production hosts. Genome streamlining has been demonstrated to be a successful approach for improving productivity and fitness in heterotrophs but is yet to be explored to its full potential in phototrophs. Here, we present the systematic reduction of the genome of the cyanobacterium exhibiting the fastest exponential growth, Synechococcuselongatus UTEX 2973. This work, the first of its kind in a photoautotroph, involved an iterative process using state-of-the-art genome-editing technology guided by experimental analysis and computational tools. CRISPRCas3 enabled large, progressive deletions of predicted dispensable regions and aided in the identification of essential genes. The large deletions were combined to obtain a strain with 55-kb genome reduction. The strains with streamlined genome showed improvement in growth (up to 23%) and productivity (by 22.7%) as compared to the wild type (WT). This streamlining strategy not only has the potential to develop cyanobacterial strains with improved growth and productivity traits but can also facilitate a better understanding of their genome-to-phenome relationships.

Original languageEnglish
JournalmBio
Volume15
Issue number3
DOIs
StatePublished - Mar 2024

Keywords

  • CRISPR-Cas3
  • cyanobacteria
  • essential gene identification
  • genome minimization
  • large progressive deletion

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