Emerging Species and Genome Editing Tools: Future Prospects in Cyanobacterial Synthetic Biology

Abstract

Recent advances in synthetic biology and an emerging algal biotechnology market have spurred a prolific increase in the availability of molecular tools for cyanobacterial research. Nevertheless, work to date has focused primarily on only a small subset of model species, which arguably limits fundamental discovery and applied research towards wider commercialisation. Here, we review the requirements for uptake of new strains, including several recently characterised fast-growing species and promising non-model species. Furthermore, we discuss the potential applications of new techniques available for transformation, genetic engineering and regulation, including an up-to-date appraisal of current Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein (CRISPR/Cas) and CRISPR interference (CRISPRi) research in cyanobacteria. We also provide an overview of several exciting molecular tools that could be ported to cyanobacteria for more advanced metabolic engineering approaches (e.g., genetic circuit design). Lastly, we introduce a forthcoming mutant library for the model species Synechocystis sp. PCC 6803 that promises to provide a further powerful resource for the cyanobacterial research community.

Current knowledge and recent advances in understanding metabolism of the model cyanobacteriumSynechocystissp. PCC 6803

“…To bypass the laborious step of mutant generation, we are developing CyanoSource, a mutant library targeting every gene in Synechocystis. Construction of the library is outlined in Gale et al [257]. Building on our transformation and Modular Cloning (MoClo) techniques [258,259], we will collaborate with United Kingdom DNA Foundries in Norwich and Edinburgh to automate the generation of a whole genome library of gene insertion plasmids (representing 3456 coding sequences (CDSs)), and will transform Synechocystis to generate the largest available collection of known and novel cyanobacterial mutant strains.…”

Cyanobacteria: Review of Current Potentials and Applications

“…These approaches facilitated the development of intricate metabolic engineering programs that aim to design strains customized for different industrial biotechnology applications. Furthermore, there is a lot of potential for applications of novel techniques like Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR) associated protein (CRISPR/Cas) and CRISPR interference (CRISPRi) research in cyanobacteria [81].…”

“…Therefore, using genetic engineering tools, the strains with high productivity need to be further studied in detail. Although these kinds of studies are at the initial phases of exploration through advanced genome editing tools, there is a lot of potential to be explored further in the near future [81]. In addition to product development, further studies must be performed on strain upgrading to attain high productivity, maintenance of growth rate, and cell potential improvement for survival in response to stressed conditions.…”

Progress and challenges in engineering cyanobacteria as chassis for light‐driven biotechnology

“…The recently announced CyanoSource barcoded mutant library for the model species Synechocystis sp. PCC 6803 (Gale et al , ) will be a valuable resource of knockout strains for the community.…”