Protocol for detecting homologous recombination activity in cancer cells by adenovirus-based functional assay

Shih-Han Huang; Chih-Ying Lee; Peter Chi

doi:10.1016/j.xpro.2024.103403

Protocol for detecting homologous recombination activity in cancer cells by adenovirus-based functional assay

STAR Protoc. 2024 Oct 19;5(4):103403. doi: 10.1016/j.xpro.2024.103403. Online ahead of print.

Authors

Shih-Han Huang¹, Chih-Ying Lee², Peter Chi³

Affiliations

¹ Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan. Electronic address: bettyh1727@gmail.com.
² Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan. Electronic address: chih-ying.lee@dkfz-heidelberg.de.
³ Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan; Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan. Electronic address: peterhchi@ntu.edu.tw.

Abstract

Cancer cells with homologous recombination deficiency (HRD) exhibit a distinctive vulnerability to poly(ADP-ribose) polymerase inhibitors (PARPis). To address the limitations of existing methodologies incapable of providing real-time insights into homologous recombination (HR) status, we present an adenovirus-based functional assay designed to quantify cellular HR activity. Here, we delineate the step-by-step procedure for producing the adenovirus harboring an HR reporter, processing primary cells, and assessing HR activity in primary ovarian cancer cells. For complete details on the use and execution of this protocol, please refer to Lee et al.¹.

Keywords: cancer; cell-based assays; clinical protocol.