Improving microalgae for biotechnology — From genetics to synthetic biology

Hlavová, Monika; Turóczy, Zoltán; Bišová, Kateřina

doi:10.1016/j.biotechadv.2015.01.009

Cited by 109 publications

(51 citation statements)

References 146 publications

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“…Physical mutagens involve different kinds of irradiation such as UV, γ, and heavy ion beams. Due to easy implementation, UV has been known to be the most widely applied physical mutagen (Hlavova et al, 2015;Yi et al, 2015). UV light has strong genotoxic effect which induces DNA damage such as the formation of cyclobutane pyrimidine dimers (CPDs) and pyrimidine pyrimidone photoproducts (64 PPs) (Ikehata and Ono, 2011).…”

Section: Mutagenesismentioning

confidence: 99%

Exploring Valuable Lipids in Diatoms

et al. 2017

Front. Mar. Sci.

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Diatoms are one major group of algae in oceans that accounts almost half of marine primary food production and have also been identified as a promising candidate for biofuel production for their high level accumulation of lipids. They have gained increasingly attention for their potential applications in pharmaceuticals, cosmetics, nutrient supplements, and biofuels. This review aims to summarize the recent advances in diatom lipid study. Chemical structures and bioactivities of different lipid classes are discussed with a focus on valuable lipids such as fatty acids, polar lipids, steroids, and oxylipins from various diatoms species. Further, current extraction and fractionation approaches are compared and recent analytical techniques and methods are also reviewed with an emphasis on lipid class composition and fatty acid profiling. Biosynthetic pathways and key catalyzing enzymes are illustrated for a better understanding of fatty acid metabolism. Past engineering attempts toward generating appropriate diatom strains for lipid production are discussed with examples using mutagenesis, environmental stimulants, and genetic modification methods. Some possible future directions and applications of diatom-derived lipids are also proposed.

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Section: Mutagenesismentioning

confidence: 99%

Exploring Valuable Lipids in Diatoms

et al. 2017

Front. Mar. Sci.

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“…On the other hand, generation of loss‐of‐function mutants by random insertional or chemical mutagenesis, despite being an excellent strategy to screen for new loci underlying a particular function, is unable to probe gene function in a direct and targeted manner; moreover, diploid organisms with unknown sexual cycles can be difficult to tackle using this approach (Parker et al ., ). These limitations have greatly hindered hypothesis‐driven testing of gene and pathway functions, and impeded rational strain development in oleaginous microalgae (Hlavova et al ., ).…”

Section: Introductionmentioning

confidence: 97%

RNAi‐based targeted gene knockdown in the model oleaginous microalgae Nannochloropsis oceanica

Wei

Wang

et al. 2017

The Plant Journal

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Microalgae are promising feedstock for renewable fuels such as biodiesel, yet development of industrial oleaginous strains has been hindered by the paucity and inefficiency of reverse genetics tools. Here we established an efficient RNAi-based targeted gene-knockdown method for Nannochloropsis spp., which are emerging model organisms for industrial microalgal oil production. The method achieved a 40-80% success rate in Nannochloropsis oceanica strain IMET1. When transcript level of one carbonic anhydrase (CA) was inhibited by 62-83% via RNAi, mutant cells exhibited photosynthetic oxygen evolution (POE) rates that were 68-100% higher than wild-type (WT) at pH 6.0, equivalent to WT at pH 8.2, yet 39-45% lower than WT at pH 9.0. Moreover, the mutant POE rates were negatively correlated with the increase of culture pH, an exact opposite of WT. Thus, a dynamic carbon concentration mechanism (CCM) that is highly sensitive to pH homeostasis was revealed, where the CA inhibition likely partially abrogated the mechanism that normally deactivates CCM under a high level of dissolved CO . Extension of the method to another sequenced N. oceanica strain of CCMP 1779 demonstrated comparable performance. Finally, McrBC-PCR followed by bisulfite sequencing revealed that the gene knockdown is mediated by the CG, CHG and CHH types of DNA methylation at the coding region of the targeted gene. The efficiency, robustness and general applicability of this reverse genetics approach suggested the possibility of large-scale RNAi-based gene function screening in industrial microalgae.

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“…More precise genome editing tools such as zinc-finger nuclease (ZFN), transcription activator-like effectors (TALEs) and clustered regularly interspaced short palindromic repeats (CRISPR) have also been developed for gene activation, deletion and replacement in organisms as emerging technology. However, only a limited number of studies have been reported on engineering microalgae through these advanced genome editing tools, for example, ZFN-mediated gene editing in the green microalga C. reinhardtii and the use of TALEs in the marine diatom P. tricornutum [54]. It was reported that using CRISPR/Cas9 system for targeted gene modification in C. reinhardtii succeeded with an expression of Cas9 and single guide RNA (sgRNA) genes while the study suggested the failure to recover transformants of C. reinhardtii was caused by the toxicity of Cas9 produced constitutively following gene editing [55].…”

Section: Approaches For Developing Algal Cell Factoriesmentioning

confidence: 99%

Algal Cell Factories: Approaches, Applications, and Potentials

Fu¹,

Chaiboonchoe²,

Khraiwesh

et al. 2016

Marine Drugs

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With the advent of modern biotechnology, microorganisms from diverse lineages have been used to produce bio-based feedstocks and bioactive compounds. Many of these compounds are currently commodities of interest, in a variety of markets and their utility warrants investigation into improving their production through strain development. In this review, we address the issue of strain improvement in a group of organisms with strong potential to be productive “cell factories”: the photosynthetic microalgae. Microalgae are a diverse group of phytoplankton, involving polyphyletic lineage such as green algae and diatoms that are commonly used in the industry. The photosynthetic microalgae have been under intense investigation recently for their ability to produce commercial compounds using only light, CO2, and basic nutrients. However, their strain improvement is still a relatively recent area of work that is under development. Importantly, it is only through appropriate engineering methods that we may see the full biotechnological potential of microalgae come to fruition. Thus, in this review, we address past and present endeavors towards the aim of creating productive algal cell factories and describe possible advantageous future directions for the field.

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Improving microalgae for biotechnology — From genetics to synthetic biology

Cited by 109 publications

References 146 publications

Exploring Valuable Lipids in Diatoms

Exploring Valuable Lipids in Diatoms

RNAi‐based targeted gene knockdown in the model oleaginous microalgae Nannochloropsis oceanica

Algal Cell Factories: Approaches, Applications, and Potentials

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