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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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Hauptmann et al. (Tue,) studied this question.
synapsesocial.com/papers/69d7f176ba18484428d1833e — DOI: https://doi.org/10.1073/pnas.1506116112
Judith Hauptmann
Silence Therapeutics (Germany)
Daniel Schraivogel
Open Targets
Astrid Bruckmann
University of Regensburg
Proceedings of the National Academy of Sciences
Howard Hughes Medical Institute
National Institutes of Health
Rockefeller University
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