Title: Methods to Study Trinucleotide Repeat Instability Induced by DNA Damage and Repair.
Authors: Lai, Yanhao; Lei, Ruipeng; Ren, Yaou; Liu, Yuan
Published In Methods Mol Biol, (2019)
Abstract: Trinucleotide repeat (TNR) instability (expansion and deletion) is associated with more than 42 human neurodegenerative diseases and cancer and mediated by DNA replication, repair, recombination, and gene transcription. Somatic TNR instability is involved in the progression of TNR expansion diseases and can be modulated by DNA damage repair and gene transcription. Recent studies from our group and others have shown that DNA base damage and its repair play an active role in modulating TNR instability and are responsible for somatic age-dependent CAG repeat expansion in neurons of Huntington's disease mice induced by oxidative DNA damage. However, it remains to be elucidated how DNA damage, non-B form DNA structures, and DNA repair enzymes and cofactors can coordinate to regulate somatic TNR instability. Understanding the molecular mechanisms underlying DNA damage and repair-mediated somatic TNR instability is critically important for identification of new therapeutic targets for treatment and prevention of TNR-related diseases. Here we describe the methods to study the locations and distribution of DNA base lesions and their effects on TNR instability through DNA base excision repair in in vitro reconstituted human systems.
PubMed ID: 31127571
MeSH Terms: DNA Damage*; DNA Repair Enzymes/metabolism; DNA Repair*; DNA/genetics; DNA/isolation & purification; DNA/metabolism; Genomics/methods*; Oligonucleotides/genetics; Oligonucleotides/isolation & purification; Oligonucleotides/metabolism; Plasmids/genetics; Polymerase Chain Reaction/methods; Sequence Deletion; Trinucleotide Repeat Expansion*