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Related Experiment Video

Updated: Jun 14, 2025

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Published on: June 17, 2015

Capture of RNA-binding proteins across mouse tissues using HARD-AP.

Yijia Ren¹, Hongyu Liao¹, Jun Yan²

¹Key Laboratory of Birth Defects and Related Disease of Women and Children of MOE, Department of Pediatrics, West China Second University Hospital, State Key Laboratory of Biotherapy, Sichuan University, Chengdu, Sichuan, 610041, China.

Nature Communications

|September 28, 2024

Summary

This summary is machine-generated.

Researchers developed HARD-AP to map RNA-binding proteins (RBPs) and their interactions across mouse organs. This powerful method reveals tissue-specific RBP roles and identifies new RNA-binding domains, advancing RNA metabolism understanding.

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Related Experiment Videos

Last Updated: Jun 14, 2025

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Published on: June 17, 2015

Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

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Published on: July 22, 2016

Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis

Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis

Published on: May 10, 2019

Area of Science:

Molecular Biology
Genomics
Proteomics

Background:

RNA-binding proteins (RBPs) are crucial regulators of RNA metabolism.
A comprehensive understanding of RBP expression and function across different tissues is currently lacking.
Existing methods may not fully capture the complexity of RNA-protein interactions in diverse biological samples.

Purpose of the Study:

To develop and validate a novel method, HARD-AP, for robustly retrieving RBPs and associated RNA regulatory complexes.
To create a comprehensive atlas of RBPs across various mouse primary organs.
To systematically map RNA-binding sites and identify novel RNA-binding domains within RBPs.

Main Methods:

Development of the HARD-AP (Highly Accurate Retrieval of Danh-associated Proteins) technique.
Application of HARD-AP to cultured cells and fresh mouse tissues.
Utilizing machine learning-based modeling to analyze and predict RNA-binding sites.
Validation of identified RBPs and RNA-binding domains, including LIM-domain-only proteins.

Main Results:

HARD-AP successfully retrieved RBPs and RNA regulatory complexes from diverse samples.
A comprehensive atlas of RBPs across mouse primary organs was established.
Machine learning modeling identified the LIM domain as a significant RNA-binding domain in numerous RBPs.
Csrp1, a LIM-domain-only protein, was validated as a tissue-dependent RNA-binding protein.

Conclusions:

HARD-AP is a powerful and versatile approach for identifying RBPomes from any sample type.
The study provides a valuable resource for understanding tissue-specific RNA-protein interactions.
This work advances the comprehensive study of RNA metabolism and regulatory networks.