CRISPR/Cas9-Based Methods for Inactivating Actinobacterial Biosynthetic Genes and Elucidating Function.

Actinobacteria / genetics CRISPR-Cas Systems / genetics Gene Editing / methods Streptomyces / genetics
Actinobacteria Biosynthetic gene CRISPR/Cas9 Natural products pCRISPomyces-2

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2022
Historique:
entrez: 6 5 2022
pubmed: 7 5 2022
medline: 11 5 2022
Statut: ppublish

Résumé

The CRISPR/Cas9 technology allows fast and marker-less genome engineering that can be employed to study secondary metabolism in actinobacteria. Here, we report a standard experimental protocol for the deletion of a biosynthetic gene in a Streptomyces species, using the vector pCRISPomyces-2 developed by Huimin Zhao and collaborators. We also describe how carrying out metabolite analysis can reveal the putative biosynthetic function of the inactivated gene.

Identifiants

DOI: 10.1007/978-1-0716-2273-5_11 PMID: 35524052
pubmed: 35524052
doi: 10.1007/978-1-0716-2273-5_11
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

201-222

Informations de copyright

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Références

Jinek M, Chylinski K, Fonfara I et al (2012) Programmable dual-RNA—guided DNA endonuclease in adaptive bacterial immunity. Science 337(6096):816–821. https://doi.org/10.1126/science.1225829
doi: 10.1126/science.1225829 pubmed: 22745249 pmcid: 6286148
Cobb RE, Wang Y, Zhao H (2015) High-efficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/Cas system. ACS Synth Biol 4(6):723–728. https://doi.org/10.1021/sb500351f
doi: 10.1021/sb500351f pubmed: 25458909
Huang H, Zheng G, Jiang W et al (2015) One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces. Acta Biochim Biophys Sin Shanghai 47(4):231–243. https://doi.org/10.1093/abbs/gmv007
doi: 10.1093/abbs/gmv007 pubmed: 25739462
Tong Y, Charusanti P, Zhang L et al (2015) CRISPR-Cas9 based engineering of Actinomycetal genomes. ACS Synth Biol 4(9):1020–1029. https://doi.org/10.1021/acssynbio.5b00038
doi: 10.1021/acssynbio.5b00038 pubmed: 25806970
Zeng H, Wen S, Xu W et al (2015) Highly efficient editing of the Actinorhodin polyketide chain length factor gene in Streptomyces coelicolor M145 using CRISPR/Cas9-CodA(sm) combined system. Appl Microbiol Biotechnol 99(24):10575–10585. https://doi.org/10.1007/s00253-015-6931-4
doi: 10.1007/s00253-015-6931-4 pubmed: 26318449
Genilloud O (2017) Actinomycetes: still a source of novel antibiotics. Nat Prod Rep 34(10):1203–1232. https://doi.org/10.1039/c7np00026j
doi: 10.1039/c7np00026j pubmed: 28820533
Alberti F, Corre C (2019) Editing Streptomycete genomes in the CRISPR/Cas9 age. Nat Prod Rep 36(9):1237–1248. https://doi.org/10.1039/C8NP00081F
doi: 10.1039/C8NP00081F pubmed: 30680376
Alberti F, Leng DJ, Wilkening I et al (2019) Triggering the expression of a silent gene cluster from genetically intractable bacteria results in Scleric acid discovery. Chem Sci 10(2):453–463. https://doi.org/10.1039/c8sc03814g
doi: 10.1039/c8sc03814g pubmed: 30746093
Blin K, Pedersen LE, Weber T et al (2016) CRISPy-web: an online resource to design sgRNAs for CRISPR applications. Synth Syst Biotechnol 1(2):118–121. https://doi.org/10.1016/j.synbio.2016.01.003
doi: 10.1016/j.synbio.2016.01.003 pubmed: 29062934 pmcid: 5640694
Kieser T, Bibb MJ, Buttner MJ et al (2000) Practical Streptomyces genetics: a laboratory manual. John Innes Foundation, Norwich
Muth G, Nußbaumer B, Wohlleben W et al (1989) A vector system with temperature-sensitive replication for gene disruption and mutational cloning in Streptomycetes. Mol Gen Genet 219(3):341–348. https://doi.org/10.1007/BF00259605
doi: 10.1007/BF00259605

Auteurs

Audam Chhun (A)

Biophore - DMF, University of Lausanne, Lausanne, Switzerland.

Fabrizio Alberti (F)

School of Life Sciences, University of Warwick, Coventry, UK. f.alberti@warwick.ac.uk.

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Classifications MeSH

questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Bâtonnets à Gram positif › Bâtonnets asporogènes à Gram positif › Bâtonnets asporogènes irréguliers à Gram positif › Actinobacteria › CRISPR/Cas9-Based Methods for Inactivating...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Systèmes CRISPR-Cas › CRISPR/Cas9-Based Methods for Inactivating...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Édition de gène › CRISPR/Cas9-Based Methods for Inactivating...
questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Bâtonnets à Gram positif › Bâtonnets sporulés à Gram positif › Streptomycetaceae › Streptomyces › CRISPR/Cas9-Based Methods for Inactivating...