Abstract
Nannochloropsis is one of the most important microalgae utilized as the feedstuff in aquaculture and is emerging model oleaginous microalgae. Recently, different genetic tools such as RNA interference, gene overexpression and gene editing have been developed in Nannochloropsis. However, the available genetic modification methods based on CRISPR technology are rather limited for modulating gene expression. Here, we firstly explored a transcriptional activation system based on CRISPR/dCas9 in N. oceanica IMET1. The result demonstrated that this construct of CRISPRa system (dCas9 protein fused with transcriptional activator VP64) could alter efficiently gene expression in N. ocanica. The expression of targeted gene (an enzyme responsible for DNA or RNA methylation as a methyltransferase) was increased by ~2–6 fold at the transcriptional level. Moreover, the growth and photosynthetic parameter (Fv/Fm) of mutants was increased by 23% and 12% compared to wild type under air-level CO2 respectively. Thus, this system will open new avenue for genome engineering of Nannochloropsis.
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