دورية أكاديمية
Chimeric RNA: DNA TracrRNA Improves Homology-Directed Repair In Vitro and In Vivo .
العنوان: | Chimeric RNA: DNA TracrRNA Improves Homology-Directed Repair In Vitro and In Vivo . |
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المؤلفون: | Simone BW; Department of Biochemistry and Molecular Biology, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Lee HB; Department of Biochemistry and Molecular Biology, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Daby CL; Department of Biochemistry and Molecular Biology, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Ata H; Department of Clinical and Translational Sciences, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Restrepo-Castillo S; Mayo Clinic Graduate School of Biomedical Sciences, Virology and Gene Therapy Track, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Martínez-Gálvez G; Mayo Clinic Graduate School of Biomedical Sciences, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Kar B; Department of Biochemistry and Molecular Biology, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Gendron WAC; Mayo Clinic Graduate School of Biomedical Sciences, Virology and Gene Therapy Track, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Clark KJ; Department of Biochemistry and Molecular Biology, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA., Ekker SC; Department of Biochemistry and Molecular Biology, Biomedical Engineering and Physiology Track, Mayo Clinic, Rochester, Minnesota, USA. |
المصدر: | The CRISPR journal [CRISPR J] 2022 Feb; Vol. 5 (1), pp. 40-52. Date of Electronic Publication: 2021 Dec 17. |
نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: Mary Ann Liebert, Inc Country of Publication: United States NLM ID: 101738191 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2573-1602 (Electronic) Linking ISSN: 25731599 NLM ISO Abbreviation: CRISPR J Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: [New Rochelle, NY] : Mary Ann Liebert, Inc., [2018]- |
مواضيع طبية MeSH: | CRISPR-Cas Systems*/genetics , Gene Editing*, DNA ; DNA Breaks, Double-Stranded ; Humans ; RNA/genetics |
مستخلص: | Nearly 90% of human pathogenic mutations are caused by small genetic variations, and methods to correct these errors efficiently are critically important. One way to make small DNA changes is providing a single-stranded oligo deoxynucleotide (ssODN) containing an alteration coupled with a targeted double-strand break (DSB) at the target locus in the genome. Coupling an ssODN donor with a CRISPR-Cas9-mediated DSB is one of the most streamlined approaches to introduce small changes. However, in many systems, this approach is inefficient and introduces imprecise repair at the genetic junctions. We herein report a technology that uses spatiotemporal localization of an ssODN with CRISPR-Cas9 to improve gene alteration. We show that by fusing an ssODN template to the trans-activating RNA (tracrRNA), we recover precise genetic alterations, with increased integration and precision in vitro and in vivo . Finally, we show that this technology can be used to enhance gene conversion with other gene editing tools such as transcription activator like effector nucleases. |
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معلومات مُعتمدة: | R01 GM134732 United States GM NIGMS NIH HHS |
المشرفين على المادة: | 63231-63-0 (RNA) 9007-49-2 (DNA) |
تواريخ الأحداث: | Date Created: 20211222 Date Completed: 20220404 Latest Revision: 20230116 |
رمز التحديث: | 20231215 |
مُعرف محوري في PubMed: | PMC8892967 |
DOI: | 10.1089/crispr.2021.0087 |
PMID: | 34935462 |
قاعدة البيانات: | MEDLINE |
تدمد: | 2573-1602 |
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DOI: | 10.1089/crispr.2021.0087 |