| Title: |
FALCON-qPCR: a new method for the quantification of oxidative lesions in mitochondrial DNA |
| Authors: |
Veronica Bazzani; Eve Harding; Szymon Balinski; Mara Equisoain Redina; Dario Alessi; Pierfrancesco Del Mestre; Walter Baratta; Daniela Cesselli; Antonio Paolo Beltrami; Michał Turek; Umberto Baccarani; Carlo Vascotto |
| Contributors: |
Bazzani, Veronica; Harding, Eve; Balinski, Szymon; Equisoain Redina, Mara; Alessi, Dario; Del Mestre, Pierfrancesco; Baratta, Walter; Cesselli, Daniela; Beltrami, Antonio Paolo; Turek, Michał; Baccarani, Umberto; Vascotto, Carlo |
| Publication Year: |
2026 |
| Collection: |
Università degli Studi di Udine: CINECA IRIS |
| Subject Terms: |
Mitochondria; mitochondrialDNA damage; oxidativestre; LongRange-PCR |
| Description: |
Accurate quantification of oxidative mitochondrial DNA (mtDNA) lesions remains technically challengingdue to the limitations of existing assays, which often require large sample inputs, multi-day workflows,and offer limited sensitivity. Here we introduce FALCON-qPCR (Fpg-assisted Long-PCR), a streamlined,high-sensitivity method for quantifying oxidative damage in mtDNA. FALCON-qPCR couples digestionwith formamidopyrimidine [fapy]-DNA glycosylase (Fpg) to long-range PCR and qPCR-based normal-ization, enabling precise lesion quantification from as few as 10,000 cells (~300 ng total DNA) withina single day. The assay provides a robust dynamic range and reproducibility across diverse biologicalsystems, including human cell lines, hepatocellular carcinoma biopsies, and Caenorhabditis elegans.Compared with established methods, FALCON-qPCR exhibits markedly higher sensitivity in detectingmtDNA damage induced by hydrogen peroxide, antimycin A, and rotenone. Its performance was furtherdemonstrated in assessing mitochondrial toxicity of ruthenium-based compounds, highlighting its poten-tial for pharmacological screening. By integrating enzymatic lesion recognition with quantitative ampli-fication in a unified workflow, FALCON-qPCR eliminates the need for mitochondrial isolation. Thismethodological advance provides a rapid, accurate, and scalable platform for studying oxidative DNAdamage, with broad applicability in mitochondrial research and translational toxicology. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| Relation: |
info:eu-repo/semantics/altIdentifier/wos/WOS:001660565300001; volume:17; issue:24; firstpage:1671; lastpage:1683; numberofpages:13; journal:BIOANALYSIS; https://hdl.handle.net/11390/1322064 |
| DOI: |
10.1080/17576180.2025.2608757 |
| Availability: |
https://hdl.handle.net/11390/1322064; https://doi.org/10.1080/17576180.2025.2608757 |
| Rights: |
info:eu-repo/semantics/openAccess ; license:Creative commons ; license uri:http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| Accession Number: |
edsbas.B4125BF2 |
| Database: |
BASE |