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Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria
Published on: January 25, 2019
Plasmid sequences and availability of a two-plasmid system for CRISPRi knockdown of Clostridioides difficile genes
Michelle Chua1,2, Daniel Erickson1,2, James Collins1,2,3
1Department of Microbiology & Immunology, University of Louisville, Louisville, Kentucky, USA.
Abstract:
A two-plasmid CRISPRi system for Clostridioides difficile that does not require antibiotic maintenance was developed. pJAK184.tetR.PT5-3.dCas9 contains an optimized tetracycline-inducible dCas9 for chromosomal insertion. pJC.15A.sgRNA.TA encodes a toxin-antitoxin system for stable maintenance, and mCherry, which is exchangeable for a customized sgRNA. We demonstrate the knockdown of the essential gene walA.
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