Identification of microRNA target genes in vivo.
MicroRNAs (miRNAs) are short non-coding RNAs transcribed from intergenic or intronic sequences as long precursors that are sequentially processed by the endonucleases Drosha and Dicer into short double-stranded sequences. It is clear that miRNAs play essential roles in gene expression, development, and cell fate specification in animals. However, one of the barriers of miRNA research is how to find the target genes. In this study, we have developed a rapid and effective method to isolate miRNA target genes in vivo. MicroRNA was synthesized in vitro and labeled by biotin. After transfected into cells, the miRNA/mRNA complexes were isolated by streptavidin-coated magnetic beads. hsa-miR155 was taken as model to validate this method, which is a very important modulator in tumor development. It is useful for validation of targets predicted in silico, and, potentially, for discovery of previously uncharacterized targets.
Zheng
Wei
W
Department Of Oncology, Shengjing Hospital of China Medical University, Sanhao Street 36, Shenyang 1100021, China. zhengwei_submit@sina.cn
Zou
Hua-Wei
HW
Tan
Yong-Gang
YG
Cai
Wei-Song
WS
D019921:Biotinylation; D045744:Cell Line, Tumor; D006801:Humans; D035683:MicroRNAs / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism*; D008954:Models, Biological; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D035683:MicroRNAs; D012333:RNA, Messenger