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TOP1 CAD-seq: A protocol to map catalytically engaged topoisomerase 1 in human cells.
Kuzin, Vladislav; Wiegard, Anika; Cameron, Donald P; Baranello, Laura.
Afiliação
  • Kuzin V; Department of Cell and Molecular Biology, Karolinska Institutet, 17177 Stockholm, Sweden. Electronic address: vladislav.kuzin@ki.se.
  • Wiegard A; Department of Cell and Molecular Biology, Karolinska Institutet, 17177 Stockholm, Sweden.
  • Cameron DP; Department of Cell and Molecular Biology, Karolinska Institutet, 17177 Stockholm, Sweden.
  • Baranello L; Department of Cell and Molecular Biology, Karolinska Institutet, 17177 Stockholm, Sweden. Electronic address: laura.baranello@ki.se.
STAR Protoc ; 3(3): 101581, 2022 09 16.
Article em En | MEDLINE | ID: mdl-35942340
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).
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA / Adutos de DNA Limite: Humans Idioma: En Revista: STAR Protoc Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA / Adutos de DNA Limite: Humans Idioma: En Revista: STAR Protoc Ano de publicação: 2022 Tipo de documento: Article