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Updated: Sep 12, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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Published on: December 11, 2020

CRISPR RiPCA for Investigating eIF4E-m⁷GpppX Capped mRNA Interactions.

Gabriela Vega-Hernández¹, Jesse Duque², Brandon J C Klein³

¹Program in Chemical Biology, University of Michigan, Ann Arbor, Michigan 48109, United States.

ACS Chemical Biology

|August 7, 2025

Summary

This summary is machine-generated.

Researchers developed CRISPR RiPCA, a new platform for studying RNA-protein interactions in live cells. This tool measures the activity of cancer drug inhibitors targeting eukaryotic translation initiation factor 4E (eIF4E) interactions.

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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

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Area of Science:

Molecular Biology
Biochemistry
Genomics

Background:

Post-transcriptional modifications of mRNA are regulated by reader RNA-binding proteins (RBPs).
Dysregulation of RBPs is linked to diseases like cancer, neurodegeneration, and viral infections.
Targeting RBPs offers a therapeutic strategy for restoring cellular function.

Purpose of the Study:

To develop a novel platform for live-cell measurement of RNA-protein interactions (RPIs).
To validate the platform using the interaction between eukaryotic translation initiation factor 4E (eIF4E) and its RNA substrate.
To demonstrate the platform's utility in assessing drug candidate efficacy.

Main Methods:

Coupling CRISPR technology with the RNA interaction with Protein-mediated Complementation Assay (RiPCA).
Establishing a live-cell RPI assay named CRISPR RiPCA.
Utilizing the eIF4E and m7G-capped RNA interaction as a model system.

Main Results:

Demonstrated the successful development of the CRISPR RiPCA platform.
Showcased the ability of CRISPR RiPCA to measure on-target activity of eIF4E inhibitors.
Validated the platform's potential for cancer drug discovery.

Conclusions:

CRISPR RiPCA is a powerful new tool for studying RPIs in live cells.
The platform enables the assessment of therapeutic interventions targeting RBPs.
This technology holds promise for advancing drug discovery in oncology and other RBP-dysregulated diseases.