In Vitro Reconstitutive Base Excision Repair (BER) Assay

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The mammalian cell genome is continuously exposed to endogenous and exogenous insults that modify its DNA. These modifications can be single-base lesions, bulky DNA adducts, base dimers, base alkylation, cytosine deamination, nitrosation, or other types of base alteration which interfere with DNA replication. Mammalian cells have evolved with a robust defense mechanism to repair these base modifications (damages) to preserve genomic stability. Base excision repair (BER) is the major defense mechanism for cells to remove these oxidative or alkylated single-base modifications. The base excision repair process involves replacement of a single-nucleotide residue by two sub-pathways, the single-nucleotide (SN) and the multi-nucleotide or long-patch (LP) base excision repair pathways. These reactions have been reproduced in vitro using cell free extracts or purified recombinant proteins involved in the base excision repair pathway. In the present chapter, we describe the detailed methodology to reconstitute base excision repair assay systems. These reconstitutive BER assay systems use artificially synthesized and modified DNA. These reconstitutive assay system will be a true representation of biologically occurring damages and their repair.

Idioma originalEnglish (US)
Título de la publicación alojadaMethods in Molecular Biology
EditorialHumana Press
Número de páginas22
EstadoPublished - 2023
Publicado de forma externa

Serie de la publicación

NombreMethods in Molecular Biology
ISSN (versión impresa)1064-3745
ISSN (versión digital)1940-6029

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology


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