Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana
Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana
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DOI：
10.1073/pnas.060034297
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发表时间：
2000-04-25
影响因子：
11.1
通讯作者：
Meyerowitz, EM
中科院分区：
文献类型：
--
作者：
Chuang, CF;Meyerowitz, EM
我们研究了双链RNA干扰（RNAi）与拟南芥基因活性的潜力。为了构建产生能够形成双链体的RNA的转化载体，将有义和反义方向的基因特异性序列连接并置于强病毒启动子的控制下。当导入A.通过农杆菌介导法转化拟南芥。对应于四个基因AGAMOUS（AG）、CLAVATA 3. APETALA 1.和PERIANTHIA，造成特定的和遗传的遗传干扰。表型的严重程度在转基因品系之间变化。原位杂交揭示了AG（RNAi）突变体中AG mRNA积累下降和表型日益严重之间的相关性，表明内源性mRNA是双链RNA介导的遗传干扰的靶点。产生稳定遗传的RNAi和由此产生的特定表型的能力使我们能够选择性地降低A. thaliana.
We investigated the potential of double-stranded RNA interference (RNAi) with gene activity in Arabidopsis thaliana. To construct transformation vectors that produce RNAs capable of duplex formation, gene-specific sequences in the sense and antisense orientations were linked and placed under the control of a strong viral promoter. When introduced into the genome of A. thaliana by Agrobacterium-mediated transformation. double-stranded RNA-expressing constructs corresponding to four genes, AGAMOUS (AG), CLAVATA3. APETALA1. and PERIANTHIA, caused specific and heritable genetic interference. The severity of phenotypes varied between transgenic lines. In situ hybridization revealed a correlation between a declining AG mRNA accumulation and increasingly severe phenotypes in AG (RNAi) mutants, suggesting that endogenous mRNA is the target of double-stranded RNA-mediated genetic interference. The ability to generate stably heritable RNAi and the resultant specific phenotypes allows us to selectively reduce gene function in A. thaliana.