Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo
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 limitati...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Public Library of Science
2014
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| Online Access: | http://hdl.handle.net/1721.1/88118 https://orcid.org/0000-0001-5891-0689 https://orcid.org/0000-0003-4698-6488 |
| _version_ | 1826211979611602944 |
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| author | Sukup-Jackson, Michelle R. Kiraly, Orsolya Na, Li Rowland, Elizabeth A. Winther, Kelly E. Chow, Danielle N. Kimoto, Takafumi Matsuguchi, Tetsuya Jonnalagadda, Vidya S. Maklakova, Vilena I. Singh, Vijay R. Wadduwage, Dushan N. Rajapakse, Jagath So, Peter T. C. Collier, Lara S. Engelward, Bevin P. Kay, Jennifer Elizabeth |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Sukup-Jackson, Michelle R. Kiraly, Orsolya Na, Li Rowland, Elizabeth A. Winther, Kelly E. Chow, Danielle N. Kimoto, Takafumi Matsuguchi, Tetsuya Jonnalagadda, Vidya S. Maklakova, Vilena I. Singh, Vijay R. Wadduwage, Dushan N. Rajapakse, Jagath So, Peter T. C. Collier, Lara S. Engelward, Bevin P. Kay, Jennifer Elizabeth |
| author_sort | Sukup-Jackson, Michelle R. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T15:14:26Z |
| format | Article |
| id | mit-1721.1/88118 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:14:26Z |
| publishDate | 2014 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/881182022-10-02T01:34:32Z Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo Sukup-Jackson, Michelle R. Kiraly, Orsolya Na, Li Rowland, Elizabeth A. Winther, Kelly E. Chow, Danielle N. Kimoto, Takafumi Matsuguchi, Tetsuya Jonnalagadda, Vidya S. Maklakova, Vilena I. Singh, Vijay R. Wadduwage, Dushan N. Rajapakse, Jagath So, Peter T. C. Collier, Lara S. Engelward, Bevin P. Kay, Jennifer Elizabeth 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. 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) 2014-06-30T13:17:36Z 2014-06-30T13:17:36Z 2014-06 2013-07 Article http://purl.org/eprint/type/JournalArticle 1553-7404 1553-7390 http://hdl.handle.net/1721.1/88118 Sukup-Jackson, Michelle R., Orsolya Kiraly, Jennifer E. Kay, Li Na, Elizabeth A. Rowland, Kelly E. Winther, Danielle N. Chow, et al. “Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo.” Edited by Peter McKinnon. PLoS Genet 10, no. 6 (June 5, 2014): e1004299. https://orcid.org/0000-0001-5891-0689 https://orcid.org/0000-0003-4698-6488 en_US http://dx.doi.org/10.1371/journal.pgen.1004299 PLoS Genetics Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Public Library of Science Public Library of Science |
| spellingShingle | Sukup-Jackson, Michelle R. Kiraly, Orsolya Na, Li Rowland, Elizabeth A. Winther, Kelly E. Chow, Danielle N. Kimoto, Takafumi Matsuguchi, Tetsuya Jonnalagadda, Vidya S. Maklakova, Vilena I. Singh, Vijay R. Wadduwage, Dushan N. Rajapakse, Jagath So, Peter T. C. Collier, Lara S. Engelward, Bevin P. Kay, Jennifer Elizabeth Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo |
| title | Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo |
| title_full | Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo |
| title_fullStr | Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo |
| title_full_unstemmed | Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo |
| title_short | Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo |
| title_sort | rosa26 gfp direct repeat radr gfp mice reveal tissue and age dependence of homologous recombination in mammals in vivo |
| url | http://hdl.handle.net/1721.1/88118 https://orcid.org/0000-0001-5891-0689 https://orcid.org/0000-0003-4698-6488 |
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