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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

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Isolation and analyses of axonal ribonucleoprotein complexes.

Ella Doron-Mandel¹, Stefanie Alber¹, Juan A Oses²

¹Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel.

Methods in Cell Biology

|January 23, 2016

Summary

This study presents methods to identify RNA-binding proteins and analyze their complexes in neuronal axons. Understanding these messenger ribonucleoprotein (mRNP) complexes is crucial for axonal function and regeneration.

Keywords:

Axon transport Proteomics RNA-binding protein RNP Sciatic nerve

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Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Published on: February 6, 2026

Area of Science:

Neuroscience
Molecular Biology
Cell Biology

Background:

Cytoskeleton-dependent RNA transport and local translation in axons are vital for neuronal maintenance and function.
Axonal transcripts regulate critical physiological processes like axon guidance, survival, and regeneration.
Messenger ribonucleoprotein (mRNP) complexes are essential for generating, maintaining, and regulating the axonal transcriptome.

Purpose of the Study:

To describe methods for studying RNA-protein interactions within neuronal axons.
To facilitate the identification of axonal RNA-binding proteins (RBPs).
To enable the analysis of messenger ribonucleoprotein (mRNP) complex dynamics.

Main Methods:

RNA affinity chromatography to identify proteins binding to a specific RNA.
Immunoprecipitation (IP) techniques to dissect protein-RNA and protein-protein interactions within mRNPs.
Analysis of mRNP dynamics under various physiological conditions.

Main Results:

Established protocols for identifying axonal RNA-binding proteins.
Demonstrated methods for analyzing protein-RNA and protein-protein interactions in mRNPs.
Provided tools to study mRNP dynamics in axons.

Conclusions:

The described methods are valuable for investigating RNA-protein interactions in axons.
These techniques advance the understanding of axonal RNA biology and mRNP complex regulation.
This work supports research into neuronal function, injury response, and regeneration.