CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers

Vogler, Brian W.; Ashford, Amy; Posewitz, Matthew C.

doi:10.1016/j.algal.2021.102385

Title: CRISPR/Cas9 disruption of glucan synthase in Nannochloropsis gaditana attenuates accumulation of β-1,3-glucose oligomers

Journal Article · Wed Jun 30 00:00:00 EDT 2021 · Algal Research

DOI:https://doi.org/10.1016/j.algal.2021.102385· OSTI ID:1843735

Vogler, Brian W. ^[1]; Ashford, Amy ^[2]; Posewitz, Matthew C. ^[2]

Colorado School of Mines, Golden, CO (United States); Conagen Inc., Bedford, MA (United States)
Colorado School of Mines, Golden, CO (United States)

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.

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Research Organization:: Colorado School of Mines, Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office; USDOE

Grant/Contract Number:: EE0007089

OSTI ID:: 1843735

Alternate ID(s):: OSTI ID: 1817269

Journal Information:: Algal Research, Vol. 58; ISSN 2211-9264

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Nannochloropsis
CRISPR/Cas9
Carbohydrate
Glucan synthase
Transglycosylase

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