Intragenic controls utilizing radiation-induced alternative transcript regions improves gene expression biodosimetry

Area of Science:

• Radiation biology
• Molecular biology
• Biotechnology

Background:

• Whole-body ionizing radiation exposure poses significant health risks, necessitating accurate biodosimetry for effective triage after radiological incidents.
• Interindividual variations in gene expression complicate precise radiation dose assessment.

Purpose of the Study:

• To develop an improved biodosimetry method for accurate radiation dose estimation.
• To address challenges posed by interindividual variations in gene expression for biodosimetry.

Main Methods:

• Lymphocyte cell lines were exposed to varying doses of ionizing radiation (0-5 Gy).
• Exon array data identified radiation-responsive alternative transcripts.
• Quantitative reverse transcription PCR (qRT-PCR) was used for gene expression analysis, employing novel cis-associated expression references for normalization.

Main Results:

• The use of cis-associated expression references improved radiation dose prediction by 2.3-8.3 fold compared to traditional normalization methods.
• This normalization strategy effectively reduced interindividual variations in gene expression.
• The method allows for estimation of basal gene expression in irradiated samples, even without pre-exposure data.

Conclusions:

• A novel normalization strategy using cis-associated expression references enhances the accuracy of gene expression-based biodosimetry.
• This approach offers a robust method to mitigate interindividual variations, improving radiation dose prediction.
• The strategy is applicable as a standalone biodosimeter or to augment existing methods, with potential broader applications in quantitative PCR.