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.
Highlights
Cyanobacteria are a diverse phylum of photosynthetic prokaryotes that are found in a wide variety of marine and freshwater habitats [1,2,3,4]
Cyanobacteria offer several advantages for biotechnological applications, including (1) higher photosynthetic efficiencies [13,14], (2) capacity to grow in hostile living environments [1,3,4], (3) the ability to be cultured on non-arable land with minimal nutrients [15], and (4) the relatively rapid and inexpensive generation of mutants [16]
Chloroplasts descend from an internalised cyanobacterium [17], certain physiological and biochemical features are conserved in eukaryotic photosynthetic organisms, making cyanobacteria excellent chassis for production of plant-derived natural products, such as terpenes [18,19]
Summary
Cyanobacteria are a diverse phylum of photosynthetic prokaryotes that are found in a wide variety of marine and freshwater habitats [1,2,3,4]. The past few years have seen a rapid proliferation of characterised tools and parts for cyanobacteria, including CRISPR/Cas-based systems [25,26,27,28] This has driven the widespread adoption of the synthetic biology paradigm for the design of biological tools based on the bottom-up approach of recombining standardised parts or modules (e.g., promoters, ribosome binding sites (RBS), coding sequences and terminators) [29,30,31,32,33,34,35]. Unmarked mutants can be generated using negative selection markers (e.g., sacB) [16,54] or by CRISPR/Cas [25]
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