Functional analysis of PIMREG deletion in DNA damage repair reporter cells

Abstract

PIMREG (PICALM interacting mitotic regulator) is a nuclear protein enriched at the nucleolus. It is a substrate of the anaphase promoter/cyclosome complex (APC/C), which controls the transition from metaphase to anaphase. We have recently shown that PIMREG is upregulated in several types of cancer, with higher expression in glioblastoma. Moreover, PIMREG expression levels increased in glioblastoma cells treated with chemotherapeutic agents or ionizing radiation suggesting that PIMREG may be related to DNA damage response. Seeking to understand the role of PIMREG in DNA damage and repair signaling pathway, we aim to knock out the PIMREG locus through the CRISPR/Cas9 system (PX458 plasmid) in a modified U2OS cell line that has a reporter system for homologous recombination (HR) DNA repair. This DNA repair reporter system will be used to identify the involvement of the PIMREG protein in the repair of DNA double-strand breaks. With a greater understanding of the role of PIMREG in DNA damage repair, it is expected to elucidate possible mechanisms of tumor cell resistance against chemotherapy and radiotherapy.(AU)