Chemical Synthesis of Lesion-Containing Oligonucleotides for DNA Repair Studies

Abstract

DNA is continuously exposed to various damaging conditions (oxidizing agents, UV, ionizing radiations, or endogenous species) that produce strand breaks, cross-links, or damaged nucleotides. In most cases, several kinds of DNA damages are simultaneously produced, often in low yield. Consequently, this hampered isolation and study of a well-defined damaged oligonucleotide (ODN). In order to get deeper insights into biological properties of a given lesion, chemistry enables preparation of large quantity of ODN containing a single and site-specifically located lesion. In addition, chemistry allows fine-tuning of lesions properties: influence of stereochemistry or of the neighboring nucleobase. This book chapter will emphasize on the methods aimed to chemical introduction of damaged nucleotides into a synthetic ODN. Phosphoramidite chemistry will be highlighted as it enables high-yielding, automatable and user-friendly site-specific incorporation of the DNA lesions.