A combinatorial system to examine the enzymatic repair of multiply damaged DNA substrates.

ABSTRACT: DNA damage drives genetic mutations that underlie the development of cancer in humans. Multiple pathways have been described in mammalian cells which can repair this damage. However, most work to date has focused upon single lesions in DNA. We present here a combinatorial system which allows assembly of duplexes containing single or multiple types of damage by ligating together six oligonucleotides containing damaged or modified bases. The combinatorial system has dual fluorescent labels allowing examination of both strands simultaneously, in order to study interactions or competition between different DNA repair pathways. Using this system, we demonstrate how repair of oxidative damage in one DNA strand can convert a mispaired T:G deamination intermediate into a T:A mutation. We also demo

SUBMITTER: Hsu CW 

PROVIDER: S-EPMC9303388 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature