RRC ID 28064
著者 Jonchere V, Bennett D.
タイトル Validating RNAi phenotypes in Drosophila using a synthetic RNAi-resistant transgene.
ジャーナル PLoS One
Abstract RNA interference (RNAi) is a powerful and widely used approach to investigate gene function, but a major limitation of the approach is the high incidence of non-specific phenotypes that arise due to off-target effects. We previously showed that RNAi-mediated knock-down of pico, which encodes the only member of the MRL family of adapter proteins in Drosophila, resulted in reduction in cell number and size leading to reduced tissue growth. In contrast, a recent study reported that pico knockdown leads to tissue dysmorphology, pointing to an indirect role for pico in the control of wing size. To understand the cause of this disparity we have utilised a synthetic RNAi-resistant transgene, which bears minimal sequence homology to the predicted dsRNA but encodes wild type Pico protein, to reanalyse the RNAi lines used in the two studies. We find that the RNAi lines from different sources exhibit different effects, with one set of lines uniquely resulting in a tissue dysmorphology phenotype when expressed in the developing wing. Importantly, the loss of tissue morphology fails to be complemented by co-overexpression of RNAi-resistant pico suggesting that this phenotype is the result of an off-target effect. This highlights the importance of careful validation of RNAi-induced phenotypes, and shows the potential of synthetic transgenes for their experimental validation.
巻・号 8(8)
ページ e70489
公開日 2013-8-8
DOI 10.1371/journal.pone.0070489
PII PONE-D-13-16039
PMID 23950943
PMC PMC3738578
MeSH Animals Base Sequence Drosophila / genetics* Male Molecular Sequence Data Phenotype RNA Interference* RNA, Small Interfering / genetics* Transgenes*
IF 2.74
引用数 8
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ショウジョウバエ 11940R-2 11940R-3