Sequencing of Captive Target Transcripts Identifies the Network of Regulated Genes and Functions of Primate-Specific miR-522
Author
Tan, Shen Mynn
Jin, Jingmin
McReynolds, Larry
Published Version
https://doi.org/10.1016/j.celrep.2014.07.023Metadata
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Tan, Shen Mynn, Rory Kirchner, Jingmin Jin, Oliver Hofmann, Larry McReynolds, Winston Hide, and Judy Lieberman. 2014. “Sequencing of Captive Target Transcripts Identifies the Network of Regulated Genes and Functions of Primate-Specific miR-522.” Cell Reports 8 (4) (August): 1225–1239. doi:10.1016/j.celrep.2014.07.023.Abstract
Identifying microRNA (miRNA)-regulated genes is key to understanding miRNA function. However, many miRNA recognition elements (MREs) do not follow canonical “seed” base-pairing rules, making identification of bona fide targets challenging. Here, we apply an unbiased sequencing-based systems approach to characterize miR-522, a member of the oncogenic primate-specific chromosome 19 miRNA cluster, highly expressed in poorly differentiated cancers. To identify miRNA targets, we sequenced full-length transcripts captured by a biotinylated miRNA mimic. Within these targets, mostly noncanonical MREs were identified by sequencing RNase-resistant fragments. miR-522 overexpression reduced mRNA, protein levels, and luciferase activity of >70% of a random list of candidate target genes and MREs. Bioinformatic analysis suggested that miR-522 regulates cell proliferation, detachment, migration, and epithelial-mesenchymal transition. miR-522 induces G1 cell-cycle arrest and causes cells to detach without anoikis, become invasive, and express mesenchymal genes. Thus, our method provides a simple but effective technique for identifying miRNA-regulated genes and biological function.Terms of Use
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http://nrs.harvard.edu/urn-3:HUL.InstRepos:26869054
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