DNA and histone proteins form chromatin, with nucleosome core particles as the smallest unit. Chromatin can be altered by histone modification or DNA damage, which is caused by cellular metabolism or by external factors such as UV and ionizing radiation, or mutagenic chemicals. Many regulatory proteins are involved in chromatin function, including non-histone proteins like HMGN1, which is an intrinsically disordered, nucleosome binding nuclear protein. However, the effects of post-translational modifications on the biological function of HMGN1, such as DNA damage, remain largely unknown. We aim to investigate the effects of lysine acetylation in the nucleosome binding domain of HMGN1 on binding affinity to damaged DNA. In this poster, we present our approach to accessing site-specifically acetylated HMGN1 variants. We examined several semi-synthetic strategies and native chemical ligation sites for their synthetic accessibility and efficiency. The site-specifically acetylated HMGN1 variants will be used for electrophoretic mobility shift assays to determine how the modified protein–damaged DNA interactions change compared to native HMGN1. Understanding how acetylation of HMGN1 modulates binding to damaged DNA and its repair by PARP1 will provide insights into the role of HMGN1 in DNA repair and packaging.