دورية أكاديمية
BE-FLARE: a fluorescent reporter of base editing activity reveals editing characteristics of APOBEC3A and APOBEC3B.
العنوان: | BE-FLARE: a fluorescent reporter of base editing activity reveals editing characteristics of APOBEC3A and APOBEC3B. |
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المؤلفون: | Coelho MA; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK. Matthew.Coelho@astrazeneca.com., Li S; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden., Pane LS; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden., Firth M; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Ciotta G; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Wrigley JD; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Cuomo ME; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Maresca M; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden., Taylor BJM; Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK. Benjamin.Taylor@astrazeneca.com. |
المصدر: | BMC biology [BMC Biol] 2018 Dec 28; Vol. 16 (1), pp. 150. Date of Electronic Publication: 2018 Dec 28. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: BioMed Central Country of Publication: England NLM ID: 101190720 Publication Model: Electronic Cited Medium: Internet ISSN: 1741-7007 (Electronic) Linking ISSN: 17417007 NLM ISO Abbreviation: BMC Biol Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: [London] : BioMed Central, c2003- |
مواضيع طبية MeSH: | APOBEC-1 Deaminase/*genetics , Cytidine Deaminase/*genetics , Gene Editing/*methods , Genetic Engineering/*methods , Minor Histocompatibility Antigens/*genetics , Proteins/*genetics, Animals ; Humans ; Rats |
مستخلص: | Background: Base Editing is a precise genome editing method that uses a deaminase-Cas9 fusion protein to mutate cytidine to thymidine in target DNA in situ without the generation of a double-strand break. However, the efficient enrichment of genetically modified cells using this technique is limited by the ability to detect such events. Results: We have developed a Base Editing FLuorescent Activity REporter (BE-FLARE), which allows for the enrichment of cells that have undergone editing of target loci based on a fluorescence shift from BFP to GFP. We used BE-FLARE to evaluate the editing efficiency of APOBEC3A and APOBEC3B family members as alternatives deaminase domains to the rat APOBEC1 domain used in base editor 3 (BE3). We identified human APOBEC3A and APOBEC3B as highly efficient cytidine deaminases for base editing applications with unique properties. Conclusions: Using BE-FLARE to report on the efficiency and precision of editing events, we outline workflows for the accelerated generation of genetically engineered cell models and the discovery of alternative base editors. |
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فهرسة مساهمة: | Keywords: APOBEC; Base editing; CRISPR/Cas9; Fluorescent reporter; Gene editing |
المشرفين على المادة: | 0 (Minor Histocompatibility Antigens) 0 (Proteins) EC 3.5.4.36 (APOBEC-1 Deaminase) EC 3.5.4.36 (Apobec1 protein, rat) EC 3.5.4.5 (APOBEC3A protein, human) EC 3.5.4.5 (APOBEC3B protein, human) EC 3.5.4.5 (Cytidine Deaminase) |
تواريخ الأحداث: | Date Created: 20181230 Date Completed: 20190503 Latest Revision: 20231005 |
رمز التحديث: | 20231215 |
مُعرف محوري في PubMed: | PMC6309101 |
DOI: | 10.1186/s12915-018-0617-1 |
PMID: | 30593278 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1741-7007 |
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DOI: | 10.1186/s12915-018-0617-1 |