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由聚合转录引发的斑马鱼中RNA干扰介导的基因沉默。

RNAi-mediated gene silencing in zebrafish triggered by convergent transcription.

作者信息

Andrews Omozusi E, Cha Diana J, Wei Chunyao, Patton James G

机构信息

1] Department of Biological Sciences, Vanderbilt University, Nashville, TN [2].

Department of Biological Sciences, Vanderbilt University, Nashville, TN.

出版信息

Sci Rep. 2014 Jun 9;4:5222. doi: 10.1038/srep05222.

Abstract

RNAi based strategies to induce gene silencing are commonly employed in numerous model organisms but have not been extensively used in zebrafish. We found that introduction of transgenes containing convergent transcription units in zebrafish embryos induced stable transcriptional gene silencing (TGS) in cis and trans for reporter (mCherry) and endogenous (One-Eyed Pinhead (OEP) and miR-27a/b) genes. Convergent transcription enabled detection of both sense and antisense transcripts and silencing was suppressed upon Dicer knockdown, indicating processing of double stranded RNA. By ChIP analyses, increased silencing was accompanied by enrichment of the heterochromatin mark H3K9me3 in the two convergently arranged promoters and in the intervening reading frame. Our work demonstrates that convergent transcription can induce gene silencing in zebrafish providing another tool to create specific temporal and spatial control of gene expression.

摘要

基于RNA干扰的基因沉默策略在众多模式生物中普遍应用,但在斑马鱼中尚未得到广泛应用。我们发现,在斑马鱼胚胎中引入含有反向转录单元的转基因,可在顺式和反式条件下对报告基因(mCherry)以及内源基因(独眼针头蛋白(OEP)和miR-27a/b)诱导稳定的转录基因沉默(TGS)。反向转录能够检测正义和反义转录本,且在Dicer敲低后沉默被抑制,这表明双链RNA得到了加工。通过染色质免疫沉淀(ChIP)分析,在两个反向排列启动子以及中间阅读框中,沉默增强伴随着异染色质标记H3K9me3的富集。我们的工作表明,反向转录可在斑马鱼中诱导基因沉默,为创建基因表达的特定时空控制提供了另一种工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/543f/4048883/604711896329/srep05222-f1.jpg

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