CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers
Abstract
Nannochloropsis species have garnered significant interest for biofuel production due to their ability to accumulate high levels of triacylglycerols (TAGs), especially following nitrogen-starvation. Furthermore, the first response to nutrient starvation is the synthesis of chrysolaminarin, a soluble β-1,3-glucan with β-1,6-branching. We employ CRISPR/Cas9 to knock out two key enzymes responsible for the synthesis of this oligosaccharide: a beta-glucan synthase (BGS) gene putatively responsible for the glucose β-1,3-linkages, and a transglycosylase (TGS) which putatively catalyzes β-1,6-branching. Analysis of the biomass from the generated mutants confirmed an ~5-fold decrease in the accumulation of soluble carbohydrate following nitrogen starvation, without an observed growth defect in a diel light-cycling regime compared to CRISPR-expressing controls.
- Authors:
-
- Colorado School of Mines, Golden, CO (United States); Conagen Inc., Bedford, MA (United States)
- Colorado School of Mines, Golden, CO (United States)
- Publication Date:
- Research Org.:
- Colorado School of Mines, Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office; USDOE
- OSTI Identifier:
- 1843735
- Alternate Identifier(s):
- OSTI ID: 1817269
- Grant/Contract Number:
- EE0007089
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Algal Research
- Additional Journal Information:
- Journal Volume: 58; Journal ID: ISSN 2211-9264
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Nannochloropsis; CRISPR/Cas9; Carbohydrate; Glucan synthase; Transglycosylase
Citation Formats
Vogler, Brian W., Ashford, Amy, and Posewitz, Matthew C. CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers. United States: N. p., 2021.
Web. doi:10.1016/j.algal.2021.102385.
Vogler, Brian W., Ashford, Amy, & Posewitz, Matthew C. CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers. United States. https://doi.org/10.1016/j.algal.2021.102385
Vogler, Brian W., Ashford, Amy, and Posewitz, Matthew C. Wed .
"CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers". United States. https://doi.org/10.1016/j.algal.2021.102385. https://www.osti.gov/servlets/purl/1843735.
@article{osti_1843735,
title = {CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers},
author = {Vogler, Brian W. and Ashford, Amy and Posewitz, Matthew C.},
abstractNote = {Nannochloropsis species have garnered significant interest for biofuel production due to their ability to accumulate high levels of triacylglycerols (TAGs), especially following nitrogen-starvation. Furthermore, the first response to nutrient starvation is the synthesis of chrysolaminarin, a soluble β-1,3-glucan with β-1,6-branching. We employ CRISPR/Cas9 to knock out two key enzymes responsible for the synthesis of this oligosaccharide: a beta-glucan synthase (BGS) gene putatively responsible for the glucose β-1,3-linkages, and a transglycosylase (TGS) which putatively catalyzes β-1,6-branching. Analysis of the biomass from the generated mutants confirmed an ~5-fold decrease in the accumulation of soluble carbohydrate following nitrogen starvation, without an observed growth defect in a diel light-cycling regime compared to CRISPR-expressing controls.},
doi = {10.1016/j.algal.2021.102385},
journal = {Algal Research},
number = ,
volume = 58,
place = {United States},
year = {Wed Jun 30 00:00:00 EDT 2021},
month = {Wed Jun 30 00:00:00 EDT 2021}
}
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