NO06G03670 knockout (KO) increased neutral lipid content without compromising growth

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 biosynthesisglycolysis 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.

Publications: