The role of DNA repair deficiency in lipid accumulation: A proof-of-concept study.

Animal models suggest an association between base excision repair (BER) deficiency and increased risk of obesity. To mechanistically investigate the effect of BER deficiency on intracellular lipid accumulation, we studied metabolic activity in in vitro BER knockdown (KD) models, targeting MutY DNA Glycosylase (MUTYH), Nth Like DNA Glycosylase 1 (NTHL1) and 8-Oxoguanine DNA Glycosylase (OGG1). We hypothesized that exposing BER deficient cells to lipids leads to reduced mitochondrial function and enhanced intracellular lipid accumulation.

An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity.

This optimized protocol (including links to instruction videos) describes a comet-based in vitro DNA repair assay that is relatively simple, versatile, and inexpensive, enabling the detection of base and nucleotide excision repair activity. Protein extracts from samples are incubated with agarose-embedded substrate nucleoids ('naked' supercoiled DNA) containing specifically induced DNA lesions (e.g.

Recent applications of the Comet Assay: A report from the International Comet Assay Workshop 2019.

The International Comet Assay Workshops (ICAW) are a series of scientific conferences dealing with different aspects of the comet assay. The assay itself is a simple method for the detection of DNA strand breaks at the cellular level and can be applied to any cell type derived from different organs and tissues of eukaryotic organisms. Additionally, the comet assay is widely applied in human biomonitoring, ecotoxicology, genotoxicity testing of chemicals, but also in basic research studying the mechanisms of DNA damage and repair.