Translation Start Sites

Exploring Translation Start Sites with Ribosome Profiling

The process of translation initiation defines the protein-coding potential of mRNAs, beginning at specific start codons, predominantly AUG. However, emerging evidence reveals that non-canonical start codons (e.g., CUG, GUG) also play critical roles in expanding proteomic diversity. The mechanisms underlying translation initiation at these sites are central to understanding gene regulation and protein synthesis. EIRNA Bio’s ribosome profiling service offers a high-resolution approach to map translation start sites across the transcriptome. This technology captures ribosome-protected mRNA fragments, pinpointing initiation codons and revealing novel regulatory elements involved in translation initiation.

Applications of Ribosome Profiling for Translation Start Site Analysis

1. Identifying Canonical and Non-Canonical Start Sites

Traditional annotation methods often overlook non-canonical translation start sites, leading to incomplete proteome characterisation. Ribosome profiling provides a genome-wide view of initiation events, distinguishing between canonical AUG codons and non-canonical sites.

Expanding Proteome Coverage: Discover novel proteoforms initiated at non-canonical codons, such as CUG, GUG, or UUG.
Uncovering Functional Diversity: Explore the roles of alternative initiation sites in cellular processes and stress responses.

2. Investigating Leaky Scanning and Reinitiation

Leaky scanning and reinitiation mechanisms allow ribosomes to bypass weak start codons or initiate translation at multiple sites within an mRNA. These processes influence the production of proteoforms with distinct functions.

Leaky Scanning: Ribosome profiling identifies start codons utilised during leaky scanning, providing insights into sequence and structural determinants of initiation efficiency.
Reinitiation Dynamics: Analyse ribosome behaviour at downstream initiation sites to uncover mechanisms regulating alternative proteoform synthesis.

3. Exploring Translational Regulation in Disease

Translation initiation is tightly regulated, and its dysregulation is implicated in various diseases, including cancer and neurodegenerative disorders. Non-canonical initiation sites often contribute to aberrant proteoform production in these conditions.

Cancer Biology: Non-canonical start sites may drive oncogenic pathways or modulate tumour suppressor activity.
Neurodegeneration: Investigate how alternative initiation contributes to proteostasis and disease progression.

4. Optimising Therapeutic Constructs

Accurate identification of translation start sites is critical for designing therapeutic gene constructs. Ribosome profiling ensures that constructs target the desired initiation codons, avoiding unintended regulatory effects.

EIRNA Bio’s Approach to Translation Start Site Analysis

High-Resolution Data for Start Site Mapping

EIRNA Bio’s ribosome profiling protocols are optimised to preserve ribosome-protected fragments, capturing initiation events at nucleotide resolution. Stringent quality control measures, including triplet periodicity analysis, ensure reliable detection of translation start sites.

Advanced Analysis with EIRNA Bio-Connect

EIRNA Bio-Connect offers robust tools for mapping and analysing translation start sites, including:

Codon-Level Resolution: Identify ribosome occupancy at canonical and non-canonical initiation codons.
Sequence Context Analysis: Characterise features influencing initiation efficiency, such as Kozak sequences.
Differential Initiation Studies: Compare initiation patterns across conditions, such as stress responses or disease states.

Collaborative Project Support

EIRNA Bio provides tailored support for translation start site studies, assisting with experimental design, data interpretation, and project-specific requirements. From discovering novel initiation mechanisms to optimising therapeutic constructs, EIRNA Bio’s expertise ensures impactful results.

Why Choose EIRNA Bio for Translation Start Site Analysis?

Translation initiation plays a fundamental role in proteome diversity and regulation. EIRNA Bio’s ribosome profiling service provides the tools and expertise needed to uncover the complexities of initiation events.