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Inhibition of pds Gene Expression via the RNA Interference Approach in Dunaliella salina (Chlorophyta)

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Abstract

To investigate the potential of double-stranded RNA interferencing with gene expression in Dunaliella salina, a plasmid pBIRNAI-Dsa was constructed to express hairpin RNA (hpRNA) containing sequences homologous to phytoene desaturase gene (pds), a key gene in carotenoid biosynthesis, and transformed into D. salina by electroporation. The relative transcription level of pds in pBIRNAI-Dsa–treated cells to nontreated cells was quantitated and the gene silencing efficiency by RNAi was evaluated via real-time polymerase chain reaction (PCR). The transcriptions of pds of the pBIRNAI-Dsa–treated group changed compared to those of the control group, and the \(2^{{ - \Delta \Delta {\text{C}}_{{\text{T}}} }} \) was lowest on the 7th day, corresponding to 0.281265-fold of the relative pds control transcript; a relatively strong gene inhibition effect was therefore deduced. The transcript of pds may be modulated in a wide range, and a reduced transcription even to 28% of the normal level may be tolerated for its survival. This study shows that dsRNA-mediated genetic interference can induce sequence-specific inhibition of gene expression and pBIRNAI-Dsa can be used for transient suppression of gene expression in D. salina. The aim of this study was to exploit dsRNA-mediated gene silencing and to provide a foundation for gene function research in D. salina.

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Acknowledgment

We thank two anonymous referees for their valuable advice and suggestions.

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Authors and Affiliations

  1. Laboratory of Marine Genetics and Breeding (MGB), College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, People’s Republic of China

    Guohua Sun, Xuecheng Zhang, Zhenghong Sui & Yunxiang Mao

  2. Marine Fisheries Research Institute of Shandong Province, Yantai, 264006, People’s Republic of China

    Guohua Sun

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  1. Guohua Sun
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  2. Xuecheng Zhang
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  3. Zhenghong Sui
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  4. Yunxiang Mao
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Correspondence to Zhenghong Sui.

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Sun, G., Zhang, X., Sui, Z. et al. Inhibition of pds Gene Expression via the RNA Interference Approach in Dunaliella salina (Chlorophyta). Mar Biotechnol 10, 219–226 (2008). https://doi.org/10.1007/s10126-007-9056-7

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  • DOI: https://doi.org/10.1007/s10126-007-9056-7

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