BIOMAT Database Logo BIOMAT Database Logo

Creating localized DNA double-strand breaks with microirradiation. 2011

Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Atomic Bomb Disease Institute, Course of Life Sciences and Radiation Research, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. kzsuzuki@nagasaki-u.ac.jp

We describe a protocol for creating localized DNA double-strand breaks (DSBs) with minimal requirements that can be applied in cell biology and molecular biology. This protocol is based on the combination of 5-bromo-2'-deoxyuridine (BrdU) labeling and ultraviolet C (UVC) irradiation through porous membranes. Cells are labeled with 10 μM BrdU for 48-72 h, washed with Ca(2+)- and Mg(2+)-free PBS(-), covered by polycarbonate membranes with micropores and exposed to UVC light. With this protocol, localized DSBs are created within subnuclear areas, irrespective of the cell cycle phase. Recruitment of proteins involved in DNA repair, DNA damage response, chromatin remodeling and histone modifications can be visualized without any specialized equipment. The quality is the same as that obtained by laser microirradiation or by any other focal irradiation. DSBs become visible within 30 min of UVC irradiation.

UI MeSH Term Description Entries
D008851 Micropore Filters A membrane or barrier with micrometer sized pores used for separation purification processes. Millipore Filters,Filter, Micropore,Filter, Millipore,Filters, Micropore,Filters, Millipore,Micropore Filter,Millipore Filter
D001973 Bromodeoxyuridine A nucleoside that substitutes for thymidine in DNA and thus acts as an antimetabolite. It causes breaks in chromosomes and has been proposed as an antiviral and antineoplastic agent. It has been given orphan drug status for use in the treatment of primary brain tumors. BUdR,BrdU,Bromouracil Deoxyriboside,Broxuridine,5-Bromo-2'-deoxyuridine,5-Bromodeoxyuridine,NSC-38297,5 Bromo 2' deoxyuridine,5 Bromodeoxyuridine,Deoxyriboside, Bromouracil
D014466 Ultraviolet Rays That portion of the electromagnetic spectrum immediately below the visible range and extending into the x-ray frequencies. The longer wavelengths (near-UV or biotic or vital rays) are necessary for the endogenous synthesis of vitamin D and are also called antirachitic rays; the shorter, ionizing wavelengths (far-UV or abiotic or extravital rays) are viricidal, bactericidal, mutagenic, and carcinogenic and are used as disinfectants. Actinic Rays,Black Light, Ultraviolet,UV Light,UV Radiation,Ultra-Violet Rays,Ultraviolet Light,Ultraviolet Radiation,Actinic Ray,Light, UV,Light, Ultraviolet,Radiation, UV,Radiation, Ultraviolet,Ray, Actinic,Ray, Ultra-Violet,Ray, Ultraviolet,Ultra Violet Rays,Ultra-Violet Ray,Ultraviolet Black Light,Ultraviolet Black Lights,Ultraviolet Radiations,Ultraviolet Ray
D016297 Mutagenesis, Site-Directed Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion. Mutagenesis, Oligonucleotide-Directed,Mutagenesis, Site-Specific,Oligonucleotide-Directed Mutagenesis,Site-Directed Mutagenesis,Site-Specific Mutagenesis,Mutageneses, Oligonucleotide-Directed,Mutageneses, Site-Directed,Mutageneses, Site-Specific,Mutagenesis, Oligonucleotide Directed,Mutagenesis, Site Directed,Mutagenesis, Site Specific,Oligonucleotide Directed Mutagenesis,Oligonucleotide-Directed Mutageneses,Site Directed Mutagenesis,Site Specific Mutagenesis,Site-Directed Mutageneses,Site-Specific Mutageneses
D053903 DNA Breaks, Double-Stranded Interruptions in the sugar-phosphate backbone of DNA, across both strands adjacently. Double-Stranded DNA Breaks,Double-Strand DNA Breaks,Double-Stranded DNA Break,Break, Double-Strand DNA,Break, Double-Stranded DNA,Breaks, Double-Strand DNA,Breaks, Double-Stranded DNA,DNA Break, Double-Strand,DNA Break, Double-Stranded,DNA Breaks, Double Stranded,DNA Breaks, Double-Strand,Double Strand DNA Breaks,Double Stranded DNA Break,Double Stranded DNA Breaks,Double-Strand DNA Break

Related Publications

Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Coping with DNA double strand breaks.
December 2010, DNA repair,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Asbestos and DNA double strand breaks.
January 1999, Cancer research,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
DNA double-strand breaks in mutagenesis.
January 1993, Environmental and molecular mutagenesis,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
DNA double strand breaks and chromosomal aberrations.
January 2010, Cytogenetic and genome research,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Sensing and repairing DNA double-strand breaks.
May 2002, Carcinogenesis,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Nucleolar responses to DNA double-strand breaks.
January 2016, Nucleic acids research,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
DNA mismatch repair-induced double-strand breaks.
January 2008, DNA repair,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
High salt and DNA double-strand breaks.
December 2011, Proceedings of the National Academy of Sciences of the United States of America,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Chromatin remodeling at DNA double-strand breaks.
March 2013, Cell,
Keiji Suzuki, and Motohiro Yamauchi, and Yasuyoshi Oka, and Masatoshi Suzuki, and Shunichi Yamashita
Checkpoint Responses to DNA Double-Strand Breaks.
June 2020, Annual review of biochemistry,
Copied contents to your clipboard!