دورية أكاديمية
أعرض في EDS

Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures.

التفاصيل البيبلوغرافية
العنوان: Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures.
المؤلفون: Doyon Y; Sangamo BioSciences, Richmond, California, USA. ydoyon@sangamo.com, Vo TD, Mendel MC, Greenberg SG, Wang J, Xia DF, Miller JC, Urnov FD, Gregory PD, Holmes MC
المصدر: Nature methods [Nat Methods] 2011 Jan; Vol. 8 (1), pp. 74-9. Date of Electronic Publication: 2010 Dec 05.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Nature Pub. Group Country of Publication: United States NLM ID: 101215604 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1548-7105 (Electronic) Linking ISSN: 15487091 NLM ISO Abbreviation: Nat Methods Subsets: MEDLINE
أسماء مطبوعة: Original Publication: New York, NY : Nature Pub. Group, c2004-
مواضيع طبية MeSH: Endonucleases/*metabolism , Zinc Fingers/*physiology, DNA/metabolism ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Dimerization ; Endonucleases/genetics ; Genome ; Saccharomyces cerevisiae/enzymology ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Zinc Fingers/genetics
مستخلص: Zinc-finger nucleases (ZFNs) drive efficient genome editing by introducing a double-strand break into the targeted gene. Cleavage is induced when two custom-designed ZFNs heterodimerize upon binding DNA to form a catalytically active nuclease complex. The importance of this dimerization event for subsequent cleavage activity has stimulated efforts to engineer the nuclease interface to prevent undesired homodimerization. Here we report the development and application of a yeast-based selection system designed to functionally interrogate the ZFN dimer interface. We identified critical residues involved in dimerization through the isolation of cold-sensitive nuclease domains. We used these residues to engineer ZFNs that have superior cleavage activity while suppressing homodimerization. The improvements were portable to orthogonal domains, allowing the concomitant and independent cleavage of two loci using two different ZFN pairs. These ZFN architectures provide a general means for obtaining highly efficient and specific genome modification.
التعليقات: Comment in: Nat Methods. 2011 Jan;8(1):53-5. (PMID: 21191373)
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المشرفين على المادة: 0 (DNA-Binding Proteins)
9007-49-2 (DNA)
EC 3.1.- (Endonucleases)
تواريخ الأحداث: Date Created: 20101207 Date Completed: 20110204 Latest Revision: 20211020
رمز التحديث: 20221213
DOI: 10.1038/nmeth.1539
PMID: 21131970
قاعدة البيانات: MEDLINE
الوصف
تدمد:1548-7105
DOI:10.1038/nmeth.1539