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Adapted qPCR methodology to detect telomere oxidative damage (TOD) associated with telomere attrition (TA) in murine tissue

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Kidd, Elliot, Meimaridou, Eirini, Walley, Andrew and Fairbrother, Una (2023) Adapted qPCR methodology to detect telomere oxidative damage (TOD) associated with telomere attrition (TA) in murine tissue. In: ISHG - Irish society of Human Genetics - 26th Meeting, 2023, 22nd September 2023, Galway, Eire.

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Abstract

Numerous Studies suggest that oxidative stress (OS) is associated with telomere attrition (TA) but they rarely directly assay telomeric oxidative damage (TOD). TOD can alter shelterin binding, induce replication fork stalling and inhibit telomerase activity. We used a previously published method 1 to measure TOD and adapted it for use in murine tissue. Our model is a C57BL/6J mouse strain and we used qPCR and a formamidopyrimidine DNA glycosylase (FPG) enzyme-based qPCR method to directly determine TOD. FPG analysis compares reaction efficiencies before and after treatment. FPG is normally involved in base excision repairs pathways, removing oxidized Purines (notably 8-Oxo Guanine). A reduction in the efficiency of the qPCR indicates the presence of 8-oxo guanine in the telomeric tract ie TOD.

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