دورية أكاديمية
Enhancing gene editing specificity by attenuating DNA cleavage kinetics.
| العنوان: | Enhancing gene editing specificity by attenuating DNA cleavage kinetics. |
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| المؤلفون: | Miller JC; Sangamo Therapeutics, Inc., Richmond, CA, USA., Patil DP; Sangamo Therapeutics, Inc., Richmond, CA, USA., Xia DF; Sangamo Therapeutics, Inc., Richmond, CA, USA., Paine CB; Sangamo Therapeutics, Inc., Richmond, CA, USA., Fauser F; Sangamo Therapeutics, Inc., Richmond, CA, USA., Richards HW; Sangamo Therapeutics, Inc., Richmond, CA, USA., Shivak DA; Sangamo Therapeutics, Inc., Richmond, CA, USA., Bendaña YR; Sangamo Therapeutics, Inc., Richmond, CA, USA., Hinkley SJ; Sangamo Therapeutics, Inc., Richmond, CA, USA., Scarlott NA; Sangamo Therapeutics, Inc., Richmond, CA, USA., Lam SC; Sangamo Therapeutics, Inc., Richmond, CA, USA., Reik A; Sangamo Therapeutics, Inc., Richmond, CA, USA., Zhou Y; Sangamo Therapeutics, Inc., Richmond, CA, USA., Paschon DE; Sangamo Therapeutics, Inc., Richmond, CA, USA., Li P; Sangamo Therapeutics, Inc., Richmond, CA, USA., Wangzor T; Sangamo Therapeutics, Inc., Richmond, CA, USA., Lee G; Sangamo Therapeutics, Inc., Richmond, CA, USA., Zhang L; Sangamo Therapeutics, Inc., Richmond, CA, USA., Rebar EJ; Sangamo Therapeutics, Inc., Richmond, CA, USA. erebar@sangamo.com. |
| المصدر: | Nature biotechnology [Nat Biotechnol] 2019 Aug; Vol. 37 (8), pp. 945-952. Date of Electronic Publication: 2019 Jul 29. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature America Publishing Country of Publication: United States NLM ID: 9604648 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1546-1696 (Electronic) Linking ISSN: 10870156 NLM ISO Abbreviation: Nat Biotechnol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: New York Ny : Nature America Publishing Original Publication: New York, NY : Nature Pub. Co., [1996- |
| مواضيع طبية MeSH: | DNA Cleavage* , T-Lymphocytes*, Gene Editing/*methods, Base Sequence ; DNA/genetics ; DNA/metabolism ; Flow Cytometry ; Hematopoietic Stem Cells ; Humans ; K562 Cells ; Protein Domains ; RNA, Messenger |
| مستخلص: | Engineered nucleases have gained broad appeal for their ability to mediate highly efficient genome editing. However the specificity of these reagents remains a concern, especially for therapeutic applications, given the potential mutagenic consequences of off-target cleavage. Here we have developed an approach for improving the specificity of zinc finger nucleases (ZFNs) that engineers the FokI catalytic domain with the aim of slowing cleavage, which should selectively reduce activity at low-affinity off-target sites. For three ZFN pairs, we engineered single-residue substitutions in the FokI domain that preserved full on-target activity but showed a reduction in off-target indels of up to 3,000-fold. By combining this approach with substitutions that reduced the affinity of zinc fingers, we developed ZFNs specific for the TRAC locus that mediated 98% knockout in T cells with no detectable off-target activity at an assay background of ~0.01%. We anticipate that this approach, and the FokI variants we report, will enable routine generation of nucleases for gene editing with no detectable off-target activity. |
| المشرفين على المادة: | 0 (RNA, Messenger) 9007-49-2 (DNA) |
| تواريخ الأحداث: | Date Created: 20190731 Date Completed: 20191106 Latest Revision: 20191115 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1038/s41587-019-0186-z |
| PMID: | 31359006 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 1546-1696 |
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| DOI: | 10.1038/s41587-019-0186-z |