A GAL4/UAS luciferase system to identify the miRNAs target mRNAs in Drosophila S2 cells.
MicroRNAs (miRNAs) have been shown to regulate gene expression through the sequence-specific base pairing with their target mRNAs. However, our understanding of the biological roles of miRNAs is still quite limited, and only a handful of miRNAs have been assigned by genetic analysis in part owing to the difficulty in the identification of their targets. Although computational methods have shown to be helpful in the prediction of miRNA targets, a major obstacle has been the lack of quick and efficient experimental procedures to verify these targets. In this report, we describe a UAS/GAL4-based reporter system for this purpose. Our data indicate it an assay of miRNA-target gene interaction, with greater sensitivity over the previously reported methods, and may be useful for more efficient identification/validation the miRNA targets in Drosophila cell lines.
Lv
Wenwen
W
Department of Genetics and Developmental Biology, Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Southeast University Medical School, 87 Dingjiaqiao Road, Nanjing, 210009, Peoples Republic of China.
Ding
Jie
J
Hu
Jiangbo
J
Wang
Chao
C
Xu
Leyi
L
Fang
Ming
M
Yuan
Liudi
L
D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins; D004330:Drosophila; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics*; D008722:Methods; D035683:MicroRNAs / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics*; D012107:Research Design; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D035683:MicroRNAs; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D008156:Luciferases