Establishing <i>Chlamydomonas reinhardtii</i> as an industrial biotechnology host

Scaife, M. A.; Nguyen, Ginnie Trinh D T; Rico, Juan José Ruiz; Lambert, Devinn; Helliwell, Katherine E.; Smith, Alison G.

doi:10.1111/tpj.12781

Cited by 176 publications

(126 citation statements)

References 151 publications

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“…Of these species, the freshwater chlorophyte Chlamydomonas reinhardtii is perhaps the most advanced microalgal platform, with a suite of molecular tools for both nuclear and chloroplast transformation, and the ongoing development of synthetic biology strategies for strain engineering [6]. The chloroplast genetic system lends itself particularly well to synthetic biology since the genome is small (205 kb) and of low complexity (99 genes) [7], and the precise integration of foreign DNA into any predetermined locus is readily achieved via homologous recombination [8].…”

Section: Introductionmentioning

confidence: 99%

The algal chloroplast as a synthetic biology platform for production of therapeutic proteins

2018

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The chloroplast of Chlamydomonas reinhardtii and other microalgae represents an attractive new platform for the synthesis of recombinant therapeutics using synthetic biology (synbio) approaches. Transgenes can be designed in silico, assembled from validated DNA parts and inserted at precise and predetermined locations within the chloroplast genome to give stable synthesis of a desired recombinant protein. Numerous recent examples of different therapeutic proteins produced successfully in the C. reinhardtii chloroplast highlight the potential of this green alga as a simple, low-cost and benign host. Furthermore, some of the features of the alga may offer additional advantages over more-established microbial, mammalian or plant-based systems. These include efficient folding and accumulation of the product in the chloroplast; a lack of contaminating toxins or infectious agents; reduced downstream processing requirements; the possibility to make complex therapeutics such as immunotoxins; and the opportunity to use the whole alga as a low-cost oral vaccine. In this paper we review the current status of algal chloroplast engineering with respect to therapeutic proteins. We also consider future advances in synbio tools, together with improvements to recipient strains, which will allow the design of bespoke strains with high levels of productivity.

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Section: Introductionmentioning

confidence: 99%

The algal chloroplast as a synthetic biology platform for production of therapeutic proteins

2018

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“…Cloning and maintenance of large DNA fragments in yeast, including the complete C. reinhardtii chloroplast genome (O'Neill et al 2012) and even entire chromosomes of Phaeodactylum tricornutum (Karas et al 2013), have already been demonstrated, and the further implementation of synthetic genomic approaches and modular functional device strategies, as illustrated by Scaife and colleagues with an example for improved triacylglycerol biosynthesis (Scaife et al 2015), are becoming realistic concepts.…”

Section: Trends and Prospectsmentioning

confidence: 98%

Genetic tools and techniques for Chlamydomonas reinhardtii

Mussgnug

2015

Appl Microbiol Biotechnol

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The development of tools has always been a major driving force for the advancement of science. Optical microscopes were the first instruments that allowed discovery and descriptive studies of the subcellular features of microorganisms. Although optical and electron microscopes remained at the forefront of microbiological research tools since their inventions, the advent of molecular genetics brought about questions which had to be addressed with new "genetic tools". The unicellular green microalgal genus Chlamydomonas, especially the most prominent species C. reinhardtii, has become a frequently used model organism for many diverse fields of research and molecular genetic analyses of C. reinhardtii, as well as the available genetic tools and techniques, have become increasingly sophisticated throughout the last decades. The aim of this review is to provide an overview of the molecular key features of C. reinhardtii and summarize the progress related to the development of tools and techniques for genetic engineering of this organism, from pioneering DNA transformation experiments to state-of-the-art techniques for targeted nuclear genome editing and high-throughput screening approaches.

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“…Many eukaryotic microalgae nuclear genomes have been transformed (Scaife et al, 2015), although only a modest number of species have been transformed and replicated by independent groups. Examples include the green algae C. reinhardtii, Volvox carteri, Chlorella ellipsoidea, Chlorella vulgaris, Dunaliella salina, and H. pluvialis; the heterokonts Phaeodactylum tricornutum, Thalassiosira pseudonana, N. gaditana and N. oceanica; and the red alga Cyanidioschyzon merolae.…”

Section: Transforming Microalgaementioning

confidence: 99%

Current advances in molecular, biochemical, and computational modeling analysis of microalgal triacylglycerol biosynthesis

Lenka

Carbonaro

Park

et al. 2016

Biotechnology Advances

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Triacylglycerols (TAGs) are highly reduced energy storage molecules ideal for biodiesel production. Microalgal TAG biosynthesis has been studied extensively in recent years, both at the molecular level and systems level through experimental studies and computational modeling. However, discussions of the strategies and products of the experimental and modeling approaches are rarely integrated and summarized together in a way that promotes collaboration among modelers and biologists in this field. In this review, we outline advances toward understanding the cellular and molecular factors regulating TAG biosynthesis in unicellular microalgae with an emphasis on recent studies on rate-limiting steps in fatty acid and TAG synthesis, while also highlighting new insights obtained from the integration of multi-omics datasets with mathematical models. Computational methodologies such as kinetic modeling, metabolic flux analysis, and new variants of flux balance analysis are explained in detail. We discuss how these methods have been used to simulate algae growth and lipid metabolism in response to changing culture conditions and how they have been used in conjunction with experimental validations. Since emerging evidence indicates that TAG synthesis in microalgae operates through coordinated crosstalk between multiple pathways in diverse subcellular destinations including the endoplasmic reticulum and plastids, we discuss new experimental studies and models that incorporate these findings for discovering key regulatory checkpoints. Finally, we describe tools for genetic manipulation of microalgae and their potential for future rational algal strain design. This comprehensive review explores the potential synergistic impact of pathway analysis, computational approaches, and molecular genetic manipulation strategies on improving TAG production in microalgae.

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Establishing Chlamydomonas reinhardtii as an industrial biotechnology host

Cited by 176 publications

References 151 publications

The algal chloroplast as a synthetic biology platform for production of therapeutic proteins

The algal chloroplast as a synthetic biology platform for production of therapeutic proteins

Genetic tools and techniques for Chlamydomonas reinhardtii

Current advances in molecular, biochemical, and computational modeling analysis of microalgal triacylglycerol biosynthesis

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