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首页> 外文期刊>Fisheries and Aquatic Sciences >Simultaneous and Systemic Knock-down of Big Defensin 1 and 2 gene Expression in the Pacific Oyster Crassostrea gigas using Long Double-stranded RNA-mediated RNA Interference
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Simultaneous and Systemic Knock-down of Big Defensin 1 and 2 gene Expression in the Pacific Oyster Crassostrea gigas using Long Double-stranded RNA-mediated RNA Interference

机译：同时使用长双链RNA介导的RNA干扰系统性地敲除太平洋牡蛎Crassostrea gigas中的大防御素1和2基因表达。

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摘要

RNA interference (RNAi)-mediated transcriptional knock-down of Crassostrea gigas big defensin 1 and 2 genes (Cg-BigDef1 and Cg-BigDef2) was investigated. The cDNA sequences of Cg-BigDef1 and Cg-BigDef2 were identical, excluding an additional fragment of 20 nucleotides in Cg-BigDef1; thus, a long double-stranded RNA (dsRNA) targeting the mRNA of Cg-BigDef2 effectively downregulated both Cg-BigDef2 and Cg-BigDef1. In addition, long dsRNA targeting green fluorescent protein (GFP) did not affect transcription of the two big defensin genes. These results suggest that the transcriptional downregulation of Cg-BigDef1 and Cg-BigDef2 was mediated by sequence-specific RNA interference (RNAi). Despite injection of long dsRNA targeting Cg-BigDef2 into only the adductor muscle, knock-down of Cg-BigDef1 and Cg-BigDef2 was observed in the adductor muscle, hemocytes, mantle, and gills, suggestive of systemic spread of RNAi in C. gigas. Furthermore, the inhibitory effect of dsRNA persisted until 72 h post-injection, indicative of a long-lasting RNAi-mediated knock-down of target genes.
机译：RNA干扰（RNAi）介导的Crassostrea gigas大防御素1和2基因（Cg-BigDef1和Cg-BigDef2）的转录敲低。 Cg-BigDef1和Cg-BigDef2的cDNA序列是相同的，除了Cg-BigDef1中另外20个核苷酸的片段。因此，靶向Cg-BigDef2 mRNA的长双链RNA（dsRNA）有效地下调了Cg-BigDef2和Cg-BigDef1。此外，靶向绿色荧光蛋白（GFP）的长dsRNA不会影响两个大防御素基因的转录。这些结果表明，Cg-BigDef1和Cg-BigDef2的转录下调是由序列特异性RNA干扰（RNAi）介导的。尽管仅向内收肌注射了靶向Cg-BigDef2的长dsRNA，但在内收肌，血细胞，地幔和g中观察到了Cg-BigDef1和Cg-BigDef2的敲低，提示RNAi在长形衣原体中的系统性传播。 。此外，dsRNA的抑制作用一直持续到注射后72小时，这表明RNAi介导的靶基因敲除作用持久。

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