Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum.
Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum.
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RNAi在豌豆蚜虫acyrthosiphon pisum中敲除基因。
DOI：
10.1186/1472-6750-7-63
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发表时间：
2007-09-28
影响因子：
3.5
通讯作者：
Tagu, Denis
中科院分区：
文献类型：
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作者：
Jaubert-Possamai, Stephanie;Le Trionnaire, Gael;Bonhomme, Joel;Christophides, Georges K;Rispe, Claude;Tagu, Denis
RNA干扰（RNAi）是一种以序列特异性方式抑制基因表达的有效方法。本研究描述了在豌豆蚜虫体内微注射双链RNA （dsRNA）的过程。将dsRNA注射到整个蚜虫体内，诱导了两种不同表达模式的标记基因的沉默：普遍表达的编码钙网蛋白的Ap-crt基因和肠道特异性的编码组织蛋白酶- l的Ap-cath-L基因。沉默的时间过程分析显示两种基因的时间模式相似：抑制在注射后1天开始，在5天达到最大，在7天停止。Ap-crt和ap -cat - l的基因表达减少了40%。豌豆蚜虫是首个基因组序列即将公布的半翅目昆虫。本研究开发的基因敲低技术将成为昆虫学进一步研究的重要后基因组工具。
RNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner. Here, we described the development of RNAi by micro-injection of double-stranded RNA (dsRNA) in the pea aphid Acyrthosiphon pisum. Injection of dsRNA into whole aphid body induced the silencing of two marker genes with different expression patterns: the ubiquitously expressed Ap-crt genes encoding a calreticulin and the gut specific Ap-cath-L gene encoding a cathepsin-L. Time-course analysis of the silencing showed similar temporal patterns for both genes: inhibition started at 1 day after injection, reached its maximum at 5 days and stopped at 7 days. A comparable 40% decrease of gene expression was observed for Ap-crt and Ap-cath-L. The pea aphid is the first Hemipteran insect for which genome sequence will be available soon. The gene knockdown technique developed in this study will be an essential post-genomic tool for further investigations in aphidology.