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Related Experiment Video

Updated: Jan 28, 2026

Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders

Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders

Published on: September 4, 2011

Structure-informed mutagenesis identifies combinatorial contributions to mouse insulin receptor IRES function.

William B Dahl¹, Tammy C T Lan², Silvi Rouskin³

¹Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02453, USA.

RNA (New York, N.Y.)

|January 26, 2026

Summary

This summary is machine-generated.

Cells use internal ribosome entry sites (IRES) to translate essential genes during stress. Researchers mapped the insulin receptor mRNA

Keywords:

5′ untranslated region RNA structure insulin receptor internal ribosome entry sites noncanonical translation initiation

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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

Area of Science:

Molecular Biology
RNA Biology
Cellular Stress Response

Background:

Cells repress cap-dependent translation during stress.
Internal ribosome entry sites (IRES) enable translation of key transcripts under stress.
The 5' untranslated region (5'UTR) of insulin receptor mRNA exhibits conserved IRES activity.

Purpose of the Study:

To investigate the RNA secondary structure of the Insr 5'UTR IRES in vitro and in cells.
To determine the structural basis of Insr IRES function.
To engineer a minimal IRES element.

Main Methods:

Dimethyl sulfate mutational profiling by sequencing (DMS-MaPseq) was used to probe RNA secondary structures.
Structural probing was performed on viral IRESes (Hepatitis C virus, Encephalomyocarditis virus) and the Insr 5'UTR IRES.
RNA structure modeling and mutational analysis were employed to identify functional RNA elements.

Main Results:

Viral IRES structures in cells were consistent with in vitro data.
A model for the mouse Insr 5'UTR IRES secondary structure was generated.
A conserved RNA segment critical for Insr IRES function was identified.
A minimal IRES element with full-length activity was designed.

Conclusions:

The study elucidates the secondary structure of a cellular IRES.
Identified key RNA elements essential for Insr IRES-mediated translation.
Developed a minimal IRES for potential therapeutic applications.