TOP1 CAD-seq: A protocol to map catalytically engaged topoisomerase 1 in human cells
Vladislav Kuzin, Anika Wiegard, Donald P. Cameron, Laura Baranello
Abstract
TOP1 CAD-seq enables mapping of TOP1 sites of covalent engagement with DNA. The procedure depends upon enrichment of DNA-covalent adducts using chaotropic salts and immunoprecipitation with an antibody specific for TOP1. Here, we describe a step-by-step protocol compatible with Illumina sequencing and bioinformatic pipeline for preliminary data analysis. Compared to other approaches for the genomic study of topoisomerases, TOP1 CAD-seq provides information about active TOP1 engaged on the DNA, taking advantage of low background due to absence of crosslinking. For complete details on the use and execution of this protocol, please refer to Das et al. (2022).
Topics & Concepts
TopoisomeraseChaotropic agentPipeline (software)Computational biologyImmunoprecipitationDNACADDNA sequencingProtocol (science)Computer scienceChemistryBiochemistryBiologyGeneProgramming languagePathologyMedicineAlternative medicineCancer therapeutics and mechanismsLung Cancer Research StudiesAdvanced biosensing and bioanalysis techniques