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Gene silencing with CRISPRi in bacteria and optimization of dCas9 expression levels.
Depardieu F, Bikard D
Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
124768pFD115pFD115 carries cas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector
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124769pFD116pFD116 carries dcas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector
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125258FD1Derivative of MG1655 used for the screening of clones able to survive the bad seed toxicity of dCas9, while still efficiently silencing the expression of mcherry
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125546pFD152derivative of pFD116 with an optimized RBS for E. coli
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