PCRISPomyces-2: Difference between revisions

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This plasmid can be used for precise CRISPR/Cas9-mediated genome engineering of <em>Streptomyces</em> strains. To our knowledge it has been successfully used in [https://doi.org/10.1128/mSphere.00305-16 S. albidoflavus], S. coelicolor, [https://pubs.rsc.org/en/content/articlelanding/2017/SC/C6SC04265A#!divAbstract S. formicae], [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4459934/ S. lividans] and S. venezuelae. Mutations have ranged from 1-100kbp deletions, precise codon changes to alter amino acids and insertions to add Flag-tags to proteins encoded at their native loci.  
This plasmid can be used for precise CRISPR/Cas9-mediated genome engineering of <em>Streptomyces</em> strains. To our knowledge it has been successfully used in [https://doi.org/10.1128/mSphere.00305-16 S. albidoflavus], S. coelicolor, [https://pubs.rsc.org/en/content/articlelanding/2017/SC/C6SC04265A#!divAbstract S. formicae], [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4459934/ S. lividans] and S. venezuelae. Mutations have ranged from 1-100kbp deletions, precise codon changes to alter amino acids and insertions to add Flag-tags to proteins encoded at their native loci.  


[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4459934/ The paper describing pCRISPomyces2]
The [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4459934/ pCRISPomyces2 paper]


The protocol from Matt Hutchings lab - [http://www.hutchingslab.uk/downloads/pCRISPOmyces2.pdf download as a PDF].
The protocol from Matt Hutchings lab - [http://www.hutchingslab.uk/downloads/pCRISPOmyces2.pdf download as a PDF].

Revision as of 20:16, 18 July 2019

This plasmid can be used for precise CRISPR/Cas9-mediated genome engineering of Streptomyces strains. To our knowledge it has been successfully used in S. albidoflavus, S. coelicolor, S. formicae, S. lividans and S. venezuelae. Mutations have ranged from 1-100kbp deletions, precise codon changes to alter amino acids and insertions to add Flag-tags to proteins encoded at their native loci.

The pCRISPomyces2 paper

The protocol from Matt Hutchings lab - download as a PDF.

The plasmid is available from AddGene under MTA: click here

CRISPR.png