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Publication : Biochemical isolation of Argonaute protein complexes by Ago-APP.

First Author  Hauptmann J Year  2015
Journal  Proc Natl Acad Sci U S A Volume  112
Issue  38 Pages  11841-5
PubMed ID  26351695 Mgi Jnum  J:226901
Mgi Id  MGI:5699185 Doi  10.1073/pnas.1506116112
Citation  Hauptmann J, et al. (2015) Biochemical isolation of Argonaute protein complexes by Ago-APP. Proc Natl Acad Sci U S A 112(38):11841-5
abstractText  During microRNA (miRNA)-guided gene silencing, Argonaute (Ago) proteins interact with a member of the TNRC6/GW protein family. Here we used a short GW protein-derived peptide fused to GST and demonstrate that it binds to Ago proteins with high affinity. This allows for the simultaneous isolation of all Ago protein complexes expressed in diverse species to identify associated proteins, small RNAs, or target mRNAs. We refer to our method as "Ago protein Affinity Purification by Peptides" (Ago-APP). Furthermore, expression of this peptide competes for endogenous TNRC6 proteins, leading to global inhibition of miRNA function in mammalian cells.
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