دورية أكاديمية
Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed That Editing Outcomes Can Vary Across Strains and Between Methods.
| العنوان: | Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed That Editing Outcomes Can Vary Across Strains and Between Methods. |
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| المؤلفون: | Leenay RT; Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695, USA., Vento JM; Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695, USA., Shah M; Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695, USA., Martino ME; Institut de Génomique Fonctionnelle de Lyon, Université de Lyon, Ecole Normale Supérieure de Lyon, Centre National de la Recherche Scientifique, Université Claude Bernard Lyon 1, Unité Mixte de Recherche 5242, 69364 Lyon, Cedex 07, France., Leulier F; Institut de Génomique Fonctionnelle de Lyon, Université de Lyon, Ecole Normale Supérieure de Lyon, Centre National de la Recherche Scientifique, Université Claude Bernard Lyon 1, Unité Mixte de Recherche 5242, 69364 Lyon, Cedex 07, France., Beisel CL; Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695, USA.; Helmholtz Institute for RNA-based Infection Research, Josef-Schneider-Straße 2, 97080, Würzburg, Germany.; Faculty of Medicine, University of Würzburg, Josef-Schneider-Straße 2, 97080, Würzburg, Germany. |
| المصدر: | Biotechnology journal [Biotechnol J] 2019 Mar; Vol. 14 (3), pp. e1700583. Date of Electronic Publication: 2018 Sep 20. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Wiley-VCH Verlag Country of Publication: Germany NLM ID: 101265833 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1860-7314 (Electronic) Linking ISSN: 18606768 NLM ISO Abbreviation: Biotechnol J Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Weinheim : Wiley-VCH Verlag, c2006- |
| مواضيع طبية MeSH: | CRISPR-Cas Systems/*genetics , Clustered Regularly Interspaced Short Palindromic Repeats/*genetics , Lactobacillus plantarum/*genetics, Gene Editing/methods ; Plasmids/genetics ; Recombinases/genetics ; Recombination, Genetic/genetics |
| مستخلص: | Lactic-acid bacteria such as Lactobacillus plantarum are commonly used for fermenting foods and as probiotics, where increasingly sophisticated genome-editing tools are employed to elucidate and enhance these microbes' beneficial properties. The most advanced tools to date utilize an oligonucleotide or double-stranded DNA donor for recombineering and Cas9 for targeted DNA cleavage. As the associated methods are often developed in isolation for one strain, it remains unclear how different Cas9-based editing methods compare across strains. Here, this work directly compares two methods in different strains of L. plantarum: one utilizing a plasmid-encoded recombineering template and another utilizing an oligonucleotide donor and an inducible DNA recombinase. This comparison reveals one instance in which only the recombineering-template method generates desired edits and another instance in which only the oligo method generates desired edits. It is further found that both methods exhibit highly variable success editing the same site across multiple L. plantarum strains. Finally, failure modes are identified for the recombineering-template method, including a consistent genomic deletion and reversion of a point mutation in the recombineering template. This study therefore highlights surprising differences for Cas9-mediated genome editing between methods and related strains, arguing for the need for multiple, distinct methods when performing CRISPR-based editing in bacteria. (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.) |
| معلومات مُعتمدة: | MCB-1452902 Division of Molecular and Cellular Biosciences; N°659510 H2020 European Research Council; FP7/2007-2013-N°309704 International European Research Council |
| فهرسة مساهمة: | Keywords: genome editing; lactobacilli; recombineering; shuttle plasmids |
| المشرفين على المادة: | 0 (Recombinases) |
| تواريخ الأحداث: | Date Created: 20180830 Date Completed: 20190614 Latest Revision: 20190614 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1002/biot.201700583 |
| PMID: | 30156038 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1860-7314 |
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| DOI: | 10.1002/biot.201700583 |