Abstract:Being the green gold of the future, microalgae and cyanobacteria have recently attracted considerable interest worldwide, for their metabolites such as lipids, protein, pigments, and bioactive compounds have immense potential for sustainable energy and pharmaceutical production capabilities. In the last decades, the efforts attended to enhance the usage of microalgae and cyanobacteria by genetic manipulation, synthetic and metabolic engineering. However, the development of photoautotrophic cell factories have … Show more
“…As a powerful toolbox for genome editing and regulation, CRISPR systems are highly valued not only for model microorganisms (e.g., E. coli, S. cerevisiae), but also provide more applicable perspectives for non-model microorganisms that are difficult to be processed through traditional methods. Despite CRISPR/Cas systems have already been applied in plenty of microbial hosts (Freed et al, 2018;Raschmanova et al, 2018;Palazzotto et al, 2019;Wang and Coleman, 2019;Ng et al, 2020), it is still challenging to construct CRISPR system with high editing efficiency, and/or apply various CRISPR strategies in non-model microorganisms. In particular, lessons have also been learned that several limitations should be overcome to enable the multiplexed /genome-scale processing of CRISPR in non-model microorganisms, such as the delivery of gRNAs or Cas proteins, the genotoxic stress, etc.…”
Section: Adaption Of Crispr/cas System To Non-model Microorganismsmentioning
The clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) system has been rapidly developed as versatile genomic engineering tools with high efficiency, accuracy and flexibility, and has revolutionized traditional methods for applications in microbial biotechnology. Here, key points of building reliable CRISPR/Cas system for genome engineering are discussed, including the Cas protein, the guide RNA and the donor DNA. Following an overview of various CRISPR/Cas tools for genome engineering, including gene activation, gene interference, orthogonal CRISPR systems and precise single base editing, we highlighted the application of CRISPR/Cas toolbox for multiplexed engineering and high throughput screening. We then summarize recent applications of CRISPR/Cas systems in metabolic engineering toward production of chemicals and natural compounds, and end with perspectives of future advancements.
“…As a powerful toolbox for genome editing and regulation, CRISPR systems are highly valued not only for model microorganisms (e.g., E. coli, S. cerevisiae), but also provide more applicable perspectives for non-model microorganisms that are difficult to be processed through traditional methods. Despite CRISPR/Cas systems have already been applied in plenty of microbial hosts (Freed et al, 2018;Raschmanova et al, 2018;Palazzotto et al, 2019;Wang and Coleman, 2019;Ng et al, 2020), it is still challenging to construct CRISPR system with high editing efficiency, and/or apply various CRISPR strategies in non-model microorganisms. In particular, lessons have also been learned that several limitations should be overcome to enable the multiplexed /genome-scale processing of CRISPR in non-model microorganisms, such as the delivery of gRNAs or Cas proteins, the genotoxic stress, etc.…”
Section: Adaption Of Crispr/cas System To Non-model Microorganismsmentioning
The clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) system has been rapidly developed as versatile genomic engineering tools with high efficiency, accuracy and flexibility, and has revolutionized traditional methods for applications in microbial biotechnology. Here, key points of building reliable CRISPR/Cas system for genome engineering are discussed, including the Cas protein, the guide RNA and the donor DNA. Following an overview of various CRISPR/Cas tools for genome engineering, including gene activation, gene interference, orthogonal CRISPR systems and precise single base editing, we highlighted the application of CRISPR/Cas toolbox for multiplexed engineering and high throughput screening. We then summarize recent applications of CRISPR/Cas systems in metabolic engineering toward production of chemicals and natural compounds, and end with perspectives of future advancements.
“…In this study, four genes were successfully edited through the expression of codon-optimized Cas9 gene and corresponding single guide RNA (sgRNA). However, the constitutive expression of Cas9 shows cytotoxic effect in C. reinhardtii that reduce the cell viability of transformants (Ng et al, 2020 ). Therefore, the transient delivery of in-vitro assembled Cas9/sg RNA ribonucleoprotein (RNP) complex via electroporation is a promising methodology to efficiently edit genes in C. reinhardtii without the cytotoxic effect of Cas9, and this approach was established recently (Shin et al, 2016a ; Baek et al, 2016a ).…”
Section: Advancement In the Resources For Microalgal Researchmentioning
Microalgae, due to their complex metabolic capacity, are being continuously explored for nutraceuticals, pharmaceuticals, and other industrially important bioactives. However, suboptimal yield and productivity of the bioactive of interest in local and robust wild-type strains are of perennial concerns for their industrial applications. To overcome such limitations, strain improvement through genetic engineering could play a decisive role. Though the advanced tools for genetic engineering have emerged at a greater pace, they still remain underused for microalgae as compared to other microorganisms. Pertaining to this, we reviewed the progress made so far in the development of molecular tools and techniques, and their deployment for microalgae strain improvement through genetic engineering. The recent availability of genome sequences and other omics datasets form diverse microalgae species have remarkable potential to guide strategic momentum in microalgae strain improvement program. This review focuses on the recent and significant improvements in the omics resources, mutant libraries, and high throughput screening methodologies helpful to augment research in the model and non-model microalgae. Authors have also summarized the case studies on genetically engineered microalgae and highlight the opportunities and challenges that are emerging from the current progress in the application of genome-editing to facilitate microalgal strain improvement. Toward the end, the regulatory and biosafety issues in the use of genetically engineered microalgae in commercial applications are described.
“…However, biofuel production from biomass of microalgal and cyanobacterial native strains (3rd generation biofuel) or their genetically modified improved strains (4th generation biofuel) has become a promising alternative to conventional biofuel production in recent years (Fig. 6 ) (Chen et al 2019a ; Mata et al 2010 ; Ng et al 2020 ; Rajneesh et al 2017 ). Fig.…”
Section: Biofuel Feedstocks Affect Food Energy and Environmentmentioning
The total number of inhabitants on the Earth is estimated to cross a record number of 9 × 10
3
million by 2050 that present a unique challenge to provide energy and clean environment to every individual. The growth in population results in a change of land use, and greenhouse gas emission due to increased industrialization and transportation. Energy consumption affects the quality of the environment by adding carbon dioxide and other pollutants to the atmosphere. This leads to oceanic acidification and other environmental fluctuations due to global climate change. Concurrently, speedy utilization of known conventional fuel reservoirs causes a challenge to a sustainable supply of energy. Therefore, an alternate energy resource is required that can maintain the sustainability of energy and environment. Among different alternatives, energy production from high carbon dioxide capturing photosynthetic aquatic microbes is an emerging technology to clean environment and produce carbon-neutral energy from their hydrocarbon-rich biomass. However, economical challenges due to low biomass production still prevent the commercialization of bioenergy. In this work, we review the impact of fossil fuels burning, which is predominantly used to fulfill global energy demand, on the quality of the environment. We also assess the status of biofuel production and utilization and discuss its potential to clean the environment. The complications associated with biofuel manufacturing using photosynthetic microorganisms are discussed and directed evolution for targeted phenotypes and targeted delivery of nutrients are proposed as potential strategies to increase the biomass production.
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