Nonsense Suppression/Readthrough

Understanding Nonsense Suppression and Readthrough with Ribosome Profiling

Protein synthesis is a highly regulated process where translation terminates when a stop codon (UAA, UAG, or UGA) is recognised in the ribosome’s A site. However, errors in termination, such as stop codon readthrough, can occur when near-cognate tRNAs outcompete termination factors. These translation failures lead to extended protein synthesis into the 3’ untranslated region (3’ UTR), which can yield truncated or malfunctioning proteins. While readthrough is often deleterious, it also presents opportunities for therapeutic development in diseases caused by premature termination codons (PTCs).

EIRNA Bio’s ribosome profiling service enables researchers to study nonsense suppression and readthrough at single-nucleotide resolution. By identifying the regulatory elements influencing translation termination, this approach provides actionable insights for understanding translational control, developing therapeutic strategies, and addressing genetic disorders.

Applications of Ribosome Profiling for Nonsense Suppression and Readthrough

1. Investigating Mechanisms of Stop Codon Readthrough

Readthrough is modulated by cis-acting elements surrounding stop codons, such as:

The stop codon identity.
Proximal nucleotides (5′ and 3′).
Stem-loop structures in the 3′ UTR.

EIRNA Bio’s ribosome profiling captures these features genome-wide, revealing the sequence and structural determinants of readthrough efficiency. This analysis helps identify the key regulatory elements involved in termination failures, advancing our understanding of translational control mechanisms.

2. Exploring Therapeutics for Premature Termination Codons

Approximately 11% of genetic disorders, such as Duchenne muscular dystrophy and cystic fibrosis, are caused by PTCs. These mutations introduce premature stop codons, truncating essential proteins. Therapeutic strategies often aim to induce selective readthrough of PTCs without affecting normal stop codons.

EIRNA Bio supports these efforts by providing high-resolution data on ribosome activity at PTC sites. By distinguishing between productive and deleterious readthrough events, researchers can refine compounds that restore protein synthesis while minimising off-target effects.

3. Analysing the Impact of Readthrough on Proteome Integrity

Unintended readthrough can disrupt cellular homeostasis by synthesising aberrant proteins or compromising proteome integrity. Ribosome profiling helps quantify readthrough rates and map their downstream effects, identifying which transcripts are affected and their consequences for cellular function.

EIRNA Bio’s service enables researchers to investigate the broader implications of translational readthrough in health and disease, supporting studies on how proteome diversity influences cellular adaptation, stress responses, and pathology.

EIRNA Bio’s Approach to Nonsense Suppression and Readthrough Analysis

High-Quality Ribosome Profiling Protocols

Ribosome profiling requires precise protocols to accurately capture ribosome-protected fragments at stop codons and 3′ UTR regions. EIRNA Bio’s methodology is designed to ensure high specificity and reproducibility, enabling reliable detection of termination failures. Quality control measures, such as triplet periodicity checks, confirm the integrity of the data, providing researchers with actionable insights into nonsense suppression.

Comprehensive Bioinformatics with EIRNA Bio-Connect

EIRNA Bio-Connect offers advanced tools for analysing readthrough events, including:

Termination Efficiency Analysis: Quantify the frequency of stop codon readthrough.
Sequence and Structural Mapping: Identify the nucleotide and structural features influencing readthrough.
Differential Analysis: Compare readthrough rates across conditions, such as treatment with readthrough-inducing compounds or disease models.

These capabilities empower researchers to explore the dynamics of nonsense suppression, offering a robust platform for understanding and addressing termination failures.

Collaborative Research Support

EIRNA Bio’s team works closely with clients to design experiments tailored to their research goals. Whether studying disease-specific readthrough events, developing therapeutic agents, or investigating translational control, EIRNA Bio provides the expertise and support needed for successful projects.

Why Choose EIRNA Bio for Readthrough Analysis?

EIRNA Bio’s ribosome profiling service combines technical excellence, advanced bioinformatics, and tailored support to provide a comprehensive solution for studying nonsense suppression and readthrough. By uncovering the regulatory elements influencing translation termination, this approach advances our understanding of translational control and its implications for disease and therapy.