التفاصيل البيبلوغرافية
| العنوان: |
Low-Toxicity and High-Efficiency Streptomyces Genome Editing Tool Based on the Miniature Type V-F CRISPR/Cas Nuclease AsCas12f1. |
| المؤلفون: |
Hua HM; College of Life Sciences, Shanghai Normal University, Shanghai 200234, China., Xu JF; College of Life Sciences, Shanghai Normal University, Shanghai 200234, China., Huang XS; Hunan Provincial Key Laboratory for Synthetic Biology of Traditional Chinese Medicine, Hunan University of Medicine, Huaihua 418000, China., Zimin AA; G.K. Scriabin Institute of Biochemistry and Physiology of Microorganisms RAS, Pushchino 142290, Russia., Wang WF; College of Life Sciences, Shanghai Normal University, Shanghai 200234, China., Lu YH; College of Life Sciences, Shanghai Normal University, Shanghai 200234, China. |
| المصدر: |
Journal of agricultural and food chemistry [J Agric Food Chem] 2024 Mar 13; Vol. 72 (10), pp. 5358-5367. Date of Electronic Publication: 2024 Mar 01. |
| نوع المنشور: |
Journal Article |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: American Chemical Society Country of Publication: United States NLM ID: 0374755 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-5118 (Electronic) Linking ISSN: 00218561 NLM ISO Abbreviation: J Agric Food Chem Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: |
Gene Editing* , Streptomyces*/genetics, CRISPR-Cas Systems ; DNA |
| مستخلص: |
Genome editing tools based on SpCas9 and FnCpf1 have facilitated strain improvements for natural product production and novel drug discovery in Streptomyces . However, due to high toxicity, their editing requires high DNA transformation efficiency, which is unavailable in most streptomycetes. The transformation efficiency of an all-in-one editing tool based on miniature Cas nuclease AsCas12f1 was significantly higher than those of SpCas9 and FnCpf1 in tested streptomycetes, which is due to its small size and weak DNA cleavage activity. Using this tool, in Streptomyces coelicolor , we achieved 100% efficiency for single gene or gene cluster deletion and 46.7 and 40% efficiency for simultaneous deletion of two genes and two gene clusters, respectively. AsCas12f1 was successfully extended to Streptomyces hygroscopicus SIPI-054 for efficient genome editing, in which SpCas9/FnCpf1 does not work well. Collectively, this work offers a low-toxicity, high-efficiency genome editing tool for streptomycetes, particularly those with low DNA transformation efficiency. |
| فهرسة مساهمة: |
Keywords: CRISPR/Cas12f; genome manipulation; industrial streptomycetes; low host toxicity; natural products |
| المشرفين على المادة: |
9007-49-2 (DNA) |
| تواريخ الأحداث: |
Date Created: 20240301 Date Completed: 20240314 Latest Revision: 20240314 |
| رمز التحديث: |
20240314 |
| DOI: |
10.1021/acs.jafc.3c09101 |
| PMID: |
38427033 |
| قاعدة البيانات: |
MEDLINE |
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