Development of CRISPR/Cas9 system in Chlorella vulgaris FSP-E to enhance lipid accumulation

Way Rong Lin, I. Son Ng

Research output: Contribution to journalArticle

Abstract

Microalgae biorefinery is an alternative, sustainable and promising trend to solve the problem of fossil oil depletion and carbon dioxide emission. However, considering the innate limitation of cell growth and oil content in microalgae, to accelerate metabolic balance by CRISPR/Cas9 system is attractive. At first, plasmid based from Agrobacterium tumefaciens and a fragment of mGFP was transformed into Chlorella sorokiniana and Chlorella vulgaris FSP-E by electroporation, respectively. Selected colonies were tested by spectrophotometer and inverted fluorescence microscopy (IFM), and an increase of fluorescent was observed by 67% compared with that in wild type, which proved the Agrobacterium-mediated plasmid is suitable for gene insertion in Chlorella species. Consequently, plasmid with similar structure as mentioned previously containing fragment of Cas9 with sgRNA designed on omega-3 fatty acid desaturase (fad3) gene was constructed and showed a higher accumulation of lipid content by 46% (w/w) in C. vulgaris FSP-E. This is first-time to use CRISPR/Cas9 based technology for gene manipulation in Chlorella.

Original languageEnglish
Article number109458
JournalEnzyme and Microbial Technology
Volume133
DOIs
Publication statusPublished - 2020 Feb

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Chlorella vulgaris
Chlorella
Lipids
Microalgae
Plasmids
Genes
Oils
Agrobacterium tumefaciens
Agrobacterium
Electroporation
Insertional Mutagenesis
Fluorescence microscopy
Spectrophotometers
Cell growth
Fluorescence Microscopy
Fatty acids
Carbon Dioxide
Carbon dioxide
Technology

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

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title = "Development of CRISPR/Cas9 system in Chlorella vulgaris FSP-E to enhance lipid accumulation",
abstract = "Microalgae biorefinery is an alternative, sustainable and promising trend to solve the problem of fossil oil depletion and carbon dioxide emission. However, considering the innate limitation of cell growth and oil content in microalgae, to accelerate metabolic balance by CRISPR/Cas9 system is attractive. At first, plasmid based from Agrobacterium tumefaciens and a fragment of mGFP was transformed into Chlorella sorokiniana and Chlorella vulgaris FSP-E by electroporation, respectively. Selected colonies were tested by spectrophotometer and inverted fluorescence microscopy (IFM), and an increase of fluorescent was observed by 67{\%} compared with that in wild type, which proved the Agrobacterium-mediated plasmid is suitable for gene insertion in Chlorella species. Consequently, plasmid with similar structure as mentioned previously containing fragment of Cas9 with sgRNA designed on omega-3 fatty acid desaturase (fad3) gene was constructed and showed a higher accumulation of lipid content by 46{\%} (w/w) in C. vulgaris FSP-E. This is first-time to use CRISPR/Cas9 based technology for gene manipulation in Chlorella.",
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Development of CRISPR/Cas9 system in Chlorella vulgaris FSP-E to enhance lipid accumulation. / Lin, Way Rong; Ng, I. Son.

In: Enzyme and Microbial Technology, Vol. 133, 109458, 02.2020.

Research output: Contribution to journalArticle

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AU - Ng, I. Son

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