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September 8, 2015Open Access

Biochemical isolation of Argonaute protein complexes by Ago-APP

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Abstract

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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Authors

Judith Hauptmann

Silence Therapeutics (Germany)

Daniel Schraivogel

Open Targets

Astrid Bruckmann

University of Regensburg

Journals

Proceedings of the National Academy of Sciences

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Howard Hughes Medical Institute

National Institutes of Health

Rockefeller University

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Hauptmann et al. (Tue,) studied this question.

synapsesocial.com/papers/69d7f176ba18484428d1833e — DOI: https://doi.org/10.1073/pnas.1506116112

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Gene Expression Omnibus: NCBI gene expression and hybridization array data repository· 2002 · 13,801 citations
The Crystal Structure of Human Argonaute2· 2012 · 704 citations

Also consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

Improved northern blot method for enhanced detection of small RNA· 2008 · 561 citations
Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencing· 2004 · 618 citations
Argonaute2 Is the Catalytic Engine of Mammalian RNAi· 2004 · 2,700 citations
Gene Expression Omnibus: NCBI gene expression and hybridization array data repository· 2002 · 13,801 citations
The Crystal Structure of Human Argonaute2· 2012 · 704 citations

Biochemical isolation of Argonaute protein complexes by Ago-APP

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Abstract

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Discussion

Authors

Journals

Actions

Institutions

References and Citations

Citation Network

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Discussion

Cite this study

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Also consider