TOP1 CAD-seq: A Protocol to Map Catalytically Engaged Topoisomerase 1 in Human Cells

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2022 Aug 9

PMID 35942340

Authors

Vladislav Kuzin

Anika Wiegard

Donald P Cameron

Laura Baranello

Affiliations

Soon will be listed here.

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).

Citing Articles

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Isolation and detection of DNA-protein crosslinks in mammalian cells.

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Coinhibition of topoisomerase 1 and BRD4-mediated pause release selectively kills pancreatic cancer via readthrough transcription.

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