In vitro base excision repair using mammalian cell extracts

Jason L Parsons; Grigory L Dianov

doi:10.1007/978-1-61779-998-3_17

In vitro base excision repair using mammalian cell extracts

Jason L Parsons, Grigory L Dianov

Cancer and Genomic Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

Base excision repair (BER) is an essential cellular mechanism that maintains genome stability by repairing DNA damage, such as DNA base lesions, base loss (AP sites) and single strand breaks, generated through endogenous metabolism or via exogenous mutagens. Therefore, in vitro BER assays are important for our understanding of the mechanism of cellular response to mutagens and may also reveal important information about the development of several DNA repair-related human diseases, such as cancer, and aging. Here, we describe the preparation and use of mammalian cell extracts in in vitro BER assays using both oligonucleotide and closed circular DNA substrates containing site-specific DNA lesions, in combination with denaturing acrylamide gel electrophoresis and phosphor imaging analysis.

Original language	English
Pages (from-to)	245-62
Number of pages	18
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	920
DOIs	https://doi.org/10.1007/978-1-61779-998-3_17
Publication status	Published - 2012

Keywords

Base Sequence
Cell Extracts/genetics
Cells, Cultured
DNA Repair
DNA Repair Enzymes/metabolism
DNA, Circular/genetics
DNA-Directed DNA Polymerase/metabolism
Denaturing Gradient Gel Electrophoresis
HeLa Cells
Humans
Molecular Imaging
Oligodeoxyribonucleotides/genetics

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/978-1-61779-998-3_17

Cite this

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title = "In vitro base excision repair using mammalian cell extracts",

abstract = "Base excision repair (BER) is an essential cellular mechanism that maintains genome stability by repairing DNA damage, such as DNA base lesions, base loss (AP sites) and single strand breaks, generated through endogenous metabolism or via exogenous mutagens. Therefore, in vitro BER assays are important for our understanding of the mechanism of cellular response to mutagens and may also reveal important information about the development of several DNA repair-related human diseases, such as cancer, and aging. Here, we describe the preparation and use of mammalian cell extracts in in vitro BER assays using both oligonucleotide and closed circular DNA substrates containing site-specific DNA lesions, in combination with denaturing acrylamide gel electrophoresis and phosphor imaging analysis.",

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year = "2012",

doi = "10.1007/978-1-61779-998-3_17",

language = "English",

volume = "920",

pages = "245--62",

journal = "Methods in molecular biology (Clifton, N.J.)",

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T1 - In vitro base excision repair using mammalian cell extracts

AU - Parsons, Jason L

AU - Dianov, Grigory L

PY - 2012

Y1 - 2012

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AB - Base excision repair (BER) is an essential cellular mechanism that maintains genome stability by repairing DNA damage, such as DNA base lesions, base loss (AP sites) and single strand breaks, generated through endogenous metabolism or via exogenous mutagens. Therefore, in vitro BER assays are important for our understanding of the mechanism of cellular response to mutagens and may also reveal important information about the development of several DNA repair-related human diseases, such as cancer, and aging. Here, we describe the preparation and use of mammalian cell extracts in in vitro BER assays using both oligonucleotide and closed circular DNA substrates containing site-specific DNA lesions, in combination with denaturing acrylamide gel electrophoresis and phosphor imaging analysis.

KW - Base Sequence

KW - Cell Extracts/genetics

KW - Cells, Cultured

KW - DNA Repair

KW - DNA Repair Enzymes/metabolism

KW - DNA, Circular/genetics

KW - DNA-Directed DNA Polymerase/metabolism

KW - Denaturing Gradient Gel Electrophoresis

KW - HeLa Cells

KW - Humans

KW - Molecular Imaging

KW - Oligodeoxyribonucleotides/genetics

U2 - 10.1007/978-1-61779-998-3_17

DO - 10.1007/978-1-61779-998-3_17

M3 - Article

C2 - 22941608

SN - 1064-3745

VL - 920

SP - 245

EP - 262

JO - Methods in molecular biology (Clifton, N.J.)

JF - Methods in molecular biology (Clifton, N.J.)

ER -

In vitro base excision repair using mammalian cell extracts

Abstract

Keywords

UN SDGs

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