Efficient genome editing in pathogenic mycobacteria using Streptococcus thermophilus CRISPR1-Cas9
| Autor: | Sergey Nejentsev, Aniek S. Meijers, Wilbert Bitter, J.J. Maaskant, Ran Troost, Coenraad Kuijl, Roy Ummels, Alexander Speer |
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| Přispěvatelé: | Molecular Microbiology, AIMMS, Medical Microbiology and Infection Prevention, Molecular cell biology and Immunology, AII - Infectious diseases |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microbiology (medical) CRISPR-Cas9 system Streptococcus thermophilus 030106 microbiology Immunology Antitubercular Agents Computational biology medicine.disease_cause Microbiology Article Mycobacterium tuberculosis 03 medical and health sciences Bacterial Proteins INDEL Mutation Genome editing SDG 3 - Good Health and Well-being CRISPR-Associated Protein 9 Drug Resistance Bacterial Isoniazid medicine CRISPR Clustered Regularly Interspaced Short Palindromic Repeats Mycobacterium marinum Gene Editing CRISPR interference biology Cas9 food and beverages Gene Expression Regulation Bacterial Catalase biology.organism_classification 3. Good health Indels 030104 developmental biology Infectious Diseases Streptococcus pyogenes CRISPR-Cas Systems Gene Deletion RNA Guide Kinetoplastida |
| Zdroj: | Tuberculosis, 124:101983. Churchill Livingstone Meijers, A S, Troost, R, Ummels, R, Maaskant, J, Speer, A, Nejentsev, S, Bitter, W & Kuijl, C P 2020, ' Efficient genome editing in pathogenic mycobacteria using Streptococcus thermophilus CRISPR1-Cas9 ', Tuberculosis, vol. 124, 101983 . https://doi.org/10.1016/j.tube.2020.101983 Tuberculosis (Edinb) Tuberculosis |
| ISSN: | 1472-9792 |
| DOI: | 10.1016/j.tube.2020.101983 |
| Popis: | The ability to genetically engineer pathogenic mycobacteria has increased significantly over the last decades due to the generation of new molecular tools. Recently, the application of the Streptococcus pyogenes and the Streptococcus thermophilus CRISPR‐Cas9 systems in mycobacteria has enabled gene editing and efficient CRISPR interference‐mediated transcriptional regulation. Here, we converted CRISPR interference into an efficient genome editing tool for mycobacteria. We demonstrate that the Streptococcus thermophilus CRISPR1-Cas9 (Sth1Cas9) is functional in Mycobacterium marinum and Mycobacterium tuberculosis, enabling highly efficient and precise DNA breaks and indel formation, without any off-target effects. In addition, with dual sgRNAs this system can be used to generate two indels simultaneously or to create specific deletions. The ability to use the power of the CRISPR-Cas9-mediated gene editing toolbox in M. tuberculosis with a single step will accelerate research into this deadly pathogen. |
| Databáze: | OpenAIRE |
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