Rosa26-GFP direct repeat (RaDR-GFP) mice reveal tissue- and age-dependence of homologous recombination in mammals in vivo

Autor: Peter T. C. So, Vilena I. Maklakova, Jagath C. Rajapakse, Tetsuya Matsuguchi, Jennifer E. Kay, Bevin P. Engelward, Michelle R. Sukup-Jackson, Li Na, Dushan N. Wadduwage, Lara S. Collier, Takafumi Kimoto, Vijay Raj Singh, Vidya S. Jonnalagadda, Elizabeth A. Rowland, Orsolya Kiraly, Kelly E. Winther, Danielle N. Chow
Přispěvatelé: Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Mechanical Engineering, Sukup-Jackson, Michelle R., Kiraly, Orsolya, Kay, Jennifer Elizabeth, Rowland, Elizabeth A., Winther, Kelly E., Chow, Danielle N., Kimoto, Takafumi, Matsuguchi, Tetsuya, Jonnalagadda, Vidya S., So, Peter T. C., Engelward, Bevin P.
Rok vydání: 2013
Předmět:
Genome instability
Cancer Research
Aging
RNA
Untranslated
DNA Repair
Liver cytology
Cancer Treatment
medicine.disease_cause
Biochemistry
Green fluorescent protein
Mice
Nucleic Acids
Molecular Cell Biology
Medicine and Health Sciences
DNA Breaks
Double-Stranded
Homologous Recombination
Genetics (clinical)
Age Factors
Brain
3. Good health
Liver
Oncology
Research Article
lcsh:QH426-470
DNA repair
DNA damage
Colon
DNA recombination
Transgene
Green Fluorescent Proteins
Mice
Transgenic
Biology
Genomic Instability
Bacterial Proteins
medicine
Genetics
Animals
Molecular Biology
Pancreas
Ecology
Evolution
Behavior and Systematics
Biology and life sciences
Cell Biology
DNA
Molecular biology
Mice
Inbred C57BL
lcsh:Genetics
Luminescent Proteins
Mutagenesis
Mutational Hypotheses
Mutation
Homologous recombination
Carcinogenesis
Zdroj: PLoS Genetics
Public Library of Science
PLoS Genetics, Vol 10, Iss 6, p e1004299 (2014)
ISSN: 1553-7404
Popis: Homologous recombination (HR) is critical for the repair of double strand breaks and broken replication forks. Although HR is mostly error free, inherent or environmental conditions that either suppress or induce HR cause genomic instability. Despite its importance in carcinogenesis, due to limitations in our ability to detect HR in vivo, little is known about HR in mammalian tissues. Here, we describe a mouse model in which a direct repeat HR substrate is targeted to the ubiquitously expressed Rosa26 locus. In the Rosa26 Direct Repeat-GFP (RaDR-GFP) mice, HR between two truncated EGFP expression cassettes can yield a fluorescent signal. In-house image analysis software provides a rapid method for quantifying recombination events within intact tissues, and the frequency of recombinant cells can be evaluated by flow cytometry. A comparison among 11 tissues shows that the frequency of recombinant cells varies by more than two orders of magnitude among tissues, wherein HR in the brain is the lowest. Additionally, de novo recombination events accumulate with age in the colon, showing that this mouse model can be used to study the impact of chronic exposures on genomic stability. Exposure to N-methyl-N-nitrosourea, an alkylating agent similar to the cancer chemotherapeutic temozolomide, shows that the colon, liver and pancreas are susceptible to DNA damage-induced HR. Finally, histological analysis of the underlying cell types reveals that pancreatic acinar cells and liver hepatocytes undergo HR and also that HR can be specifically detected in colonic somatic stem cells. Taken together, the RaDR-GFP mouse model provides new understanding of how tissue and age impact susceptibility to HR, and enables future studies of genetic, environmental and physiological factors that modulate HR in mammals.
National Institutes of Health (U.S.) (Program Project Grant P01-CA026731)
National Institutes of Health (U.S.) (R33-CA112151)
National Institute of Environmental Health Sciences (P30-ES002109)
Singapore-MIT Alliance for Research and Technology Center
National Institutes of Health (U.S.) (P41-EB015871)
National Cancer Institute (U.S.) (P30-CA014051)
Databáze: OpenAIRE
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