دورية أكاديمية
أعرض في EDS

Large-scale preparation of fluorescence multiplex host cell reactivation (FM-HCR) reporters.

التفاصيل البيبلوغرافية
العنوان: Large-scale preparation of fluorescence multiplex host cell reactivation (FM-HCR) reporters.
المؤلفون: Piett CG; Department of Environmental Health, JBL Center for Radiation Sciences, Harvard T. H. Chan School of Public Health, Boston, MA, USA., Pecen TJ; Department of Environmental Health, JBL Center for Radiation Sciences, Harvard T. H. Chan School of Public Health, Boston, MA, USA., Laverty DJ; Department of Environmental Health, JBL Center for Radiation Sciences, Harvard T. H. Chan School of Public Health, Boston, MA, USA., Nagel ZD; Department of Environmental Health, JBL Center for Radiation Sciences, Harvard T. H. Chan School of Public Health, Boston, MA, USA. znagel@hsph.harvard.edu.
المصدر: Nature protocols [Nat Protoc] 2021 Sep; Vol. 16 (9), pp. 4265-4298. Date of Electronic Publication: 2021 Aug 06.
نوع المنشور: Evaluation Study; Journal Article; Research Support, N.I.H., Extramural
اللغة: English
بيانات الدورية: Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101284307 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1750-2799 (Electronic) Linking ISSN: 17502799 NLM ISO Abbreviation: Nat Protoc Subsets: MEDLINE
أسماء مطبوعة: Original Publication: London, UK : Nature Pub. Group, 2006-
مواضيع طبية MeSH: DNA Repair* , Genes, Reporter* , Genetic Techniques*, Plasmids/*chemical synthesis, Humans
مستخلص: Repair of DNA damage is a critical survival mechanism that affects susceptibility to various human diseases and represents a key target for cancer therapy. A major barrier to applying this knowledge in research and clinical translation has been the lack of efficient, quantitative functional assays for measuring DNA repair capacity in living primary cells. To overcome this barrier, we recently developed a technology termed 'fluorescence multiplex host cell reactivation' (FM-HCR). We describe a method for using standard molecular biology techniques to generate large quantities of FM-HCR reporter plasmids containing site-specific DNA lesions and using these reporters to assess DNA repair capacity in at least six major DNA repair pathways in live cells. We improve upon previous methodologies by (i) providing a universal workflow for generating reporter plasmids, (ii) improving yield and purity to enable large-scale studies that demand milligram quantities and (iii) reducing preparation time >ten-fold.
(© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)
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معلومات مُعتمدة: P01 CA092584 United States CA NCI NIH HHS; P30 ES000002 United States ES NIEHS NIH HHS; U01 ES029520 United States ES NIEHS NIH HHS
تواريخ الأحداث: Date Created: 20210807 Date Completed: 20211013 Latest Revision: 20220716
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC9272811
DOI: 10.1038/s41596-021-00577-3
PMID: 34363069
قاعدة البيانات: MEDLINE
الوصف
تدمد:1750-2799
DOI:10.1038/s41596-021-00577-3