Repositorium
Lipid production in Nannochloropsis gaditana is doubled by decreasing expression of a single transcriptional regulator
404
Journal Article / 2017
Ajjawi, Imad; Verruto, John; Aqui, Moena; Soriaga, Leah B.; Coppersmith, Jennifer; Kwok, Kathleen; Peach, Luke; Orchard, Elizabeth; Kalb, Ryan; Xu, Weidong; Carlson, Tom J.; Francis, Kristie; Konigsfeld, Katie; Bartalis, Judit; Schultz, Andrew; Lambert, William; Schwartz, Ariel S.; Brown, Robert; Moellering, Eric R.
Lipid production in the industrial microalga Nannochloropsis gaditana exceeds that of model algal species and can be maximized by nutrient starvation in batch culture. However, starvation halts growth, thereby decreasing productivity. Efforts to engineer N. gaditana strains that can accumulate biomass and overproduce lipids have previously met with little success. We identified 20 transcription factors as putative negative regulators of lipid production by using RNA-seq analysis of N. gaditana during nitrogen deprivation. Application of a CRISPR-Cas9 reverse-genetics pipeline enabled insertional mutagenesis of 18 of these 20 transcription factors. Knocking out a homolog of fungal Zn(II)2Cys6-encoding genes improved partitioning of total carbon to lipids from 20% (wild type) to 40-55% (mutant) in nutrient-replete conditions. Knockout mutants grew poorly, but attenuation of Zn(II)2Cys6 expression yielded strains producing twice as much lipid (∼5.0 g m-2 d-1) as that in the wild type (∼2.5 g m-2 d-1) under semicontinuous growth conditions and had little effect on growth.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
928 |
Ajjawi, Imad; Moellering Eric. M USA |
Nannochloropsis gaditana |
CRISPR/Cas9 Zn2CYS |
Producing the double amount of lipids SDN3 |
Basic research Basic research |