Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA.
Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA.
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通过双链RNA的腹腔内注射，成年蜜蜂中卵黄蛋白基因功能的破坏。
DOI：
10.1186/1472-6750-3-1
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发表时间：
2003-01-20
影响因子：
3.5
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操纵成年工蜂（Apis mellifera）生理和行为的遗传网络的能力可能会加深我们对学习和记忆生成、衰老以及社会系统的调节解剖等问题的理解。在这里，我们评估了两种通过 RNA 干扰 (RNAi) 探测成年蜜蜂基因功能的方法。选择卵黄蛋白原基因作为靶标是因为其表达在蜜蜂成年阶段之前不太可能产生表型效应。这使得我们能够在前胚层卵中引入 dsRNA，而不会影响发育过程中的基因功能。在注射了源自卵黄蛋白原编码序列 504 bp 序列的 dsRNA 的卵中饲养的工蜂中，15% 的卵黄蛋白原 mRNA 水平大幅降低。当通过腹内注射将 dsRNA 引入新出现的蜜蜂时，几乎所有个体 (96%) 都表现出突变表型。 15 天后，该组中仍然存在表观大小与模板 dsRNA 相似的 RNA 片段。在前胚层阶段的卵中注射 dsRNA 似乎可以破坏所有发育阶段的基因功能。为了剖析成虫阶段的基因功能，腹内注射技术似乎优于卵注射，因为它具有更高的外显率，更简单，并且可以处理在胚胎、幼虫或蛹阶段也表达的基因。
The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. Here we assess two methods for probing gene function by RNA interference (RNAi) in adult honeybees. The vitellogenin gene was chosen as target because its expression is unlikely to have a phenotypic effect until the adult stage in bees. This allowed us to introduce dsRNA in preblastoderm eggs without affecting gene function during development. Of workers reared from eggs injected with dsRNA derived from a 504 bp stretch of the vitellogenin coding sequence, 15% had strongly reduced levels of vitellogenin mRNA. When dsRNA was introduced by intra-abdominal injection in newly emerged bees, almost all individuals (96 %) showed the mutant phenotype. An RNA-fragment with an apparent size similar to the template dsRNA was still present in this group after 15 days. Injection of dsRNA in eggs at the preblastoderm stage seems to allow disruption of gene function in all developmental stages. To dissect gene function in the adult stage, the intra-abdominal injection technique seems superior to egg injection as it gives a much higher penetrance, it is much simpler, and it makes it possible to address genes that are also expressed in the embryonic, larval or pupal stages.