Efficient and Reliable Production of Vectors for the Study of the Repair, Mutagenesis, and Phenotypic Consequences of Defined DNA Damage Lesions in Mammalian Cells
Publication Date
June 30, 2016
Journal
PLOS ONE
Authors
Lucy Petrova, Christine Gran, Magnar Bjoras & Paul W. Doetsch
Volume
11
Issue
6
Pages
e0158581
DOI
https://dx.plos.org/10.1371/journal.pone.0158581
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0158581
Scopus
84977074294
Mendeley
http://www.mendeley.com/research/efficient-reliable-production-vectors-study-repair-mutagenesis-phenotypic-consequences-defined-dna-d
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Mendeley | Further Information

{"title"=>"Efficient and Reliable Production of Vectors for the Study of the Repair, Mutagenesis, and Phenotypic Consequences of Defined DNA Damage Lesions in Mammalian Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Lucy", "last_name"=>"Petrova", "scopus_author_id"=>"55834364500"}, {"first_name"=>"Christine", "last_name"=>"Gran", "scopus_author_id"=>"57192307206"}, {"first_name"=>"Magnar", "last_name"=>"Bjoras", "scopus_author_id"=>"6603694044"}, {"first_name"=>"Paul W.", "last_name"=>"Doetsch", "scopus_author_id"=>"7005860391"}], "year"=>2016, "source"=>"PLoS ONE", "identifiers"=>{"doi"=>"10.1371/journal.pone.0158581", "scopus"=>"2-s2.0-84977074294", "issn"=>"19326203", "pui"=>"611071346", "sgr"=>"84977074294"}, "id"=>"5dff3a37-cd81-309b-a9aa-08cad3c466dd", "abstract"=>"Mammalian cells are constantly and unavoidably exposed to DNA damage from endoge-nous and exogenous sources, frequently to the detriment of genomic integrity and biological function. Cells acquire a large number of chemically diverse lesions per day, and each can have a different genetic fate and biological consequences. However, our knowledge of how and when specific lesions are repaired or how they may compromise the fidelity of DNA rep-lication or transcription and lead to deleterious biological endpoints in mammalian cells is limited. Studying individual lesions requires technically challenging approaches for the tar-geted introduction of defined lesions into relevant DNA sequences of interest. Here, we present a systematic analysis of factors influencing yield and an improved, efficient and reli-able protocol for the production of mammalian expression phagemid vectors containing defined DNA base modifications in any sequence position of either complementary DNA strand. We applied our improved protocol to study the transcriptional mutagenesis-medi-ated phenotypic consequences of the common oxidative lesion 5-hydroxyuracil, placed in the G12 mutational hotspot of the KRAS oncogene. 5-OHU induced sustained oncogenic signaling in Neil1 -/-Neil2 -/-mouse cells. The resulting advance in technology will have broad applicability for investigation of single lesion DNA repair, mutagenesis, and DNA damage responses in mammalian cells.", "link"=>"http://www.mendeley.com/research/efficient-reliable-production-vectors-study-repair-mutagenesis-phenotypic-consequences-defined-dna-d", "reader_count"=>2, "reader_count_by_academic_status"=>{"Student > Ph. D. Student"=>1, "Student > Master"=>1}, "reader_count_by_user_role"=>{"Student > Ph. D. Student"=>1, "Student > Master"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "reader_count_by_subdiscipline"=>{"Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}}, "group_count"=>0}

Scopus | Further Information

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