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Overview
Mutinfo:NO06G03670 knockout (KO) increased neutral lipid content without compromising growth |
Organism:Nannochloropsis oceanica (N. oceanica IMET1) |
Method:CRISPR/Cas |
Genes:NO06G03670 |
Selection:Staining of intracellular neutral lipids with BODIPY then FACS sorting |
Phenotypes:We constructed an insertional mutagenesis library of N. oceanica, selected high lipid mutants by five rounds of fluorescence-activated cell sorting, and identified disrupted genes using a novel implementation of a rapid genotyping procedure. One particularly promising mutant (HLM23) was disrupted in a putative APETALA2-like transcription factor gene. HLM23 showed a 40%-increased neutral lipid content, increased photosynthetic performance, and no growth impairment. Furthermore, transcriptome analysis revealed an upregulation of genes related to plastidial fatty acid biosynthesis, glycolysis and the Calvin-Benson-Bassham cycle in HLM23. Insights gained in this work can be used in future genetic engineering strategies for increased lipid productivity of Nannochloropsis.
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