Figure 6
sgRNA targeting the coding strand of dnaK gene prevents leptospiral growth in plates. L. biflexa cells were transformed with the plasmid pMaOri.dCas9 alone or containing sgRNA targeting the coding (sgRNAdnaK1) or template (sgRNAdnaK2) strand of the dnaK gene. Colonies were counted (A) and DnaK protein levels were evaluated by western blotting probing cell extracts with anti-DnaK (B, upper panel). Coomassie blue staining (B, lower panel) was used as loading control. (C) Relative transcription level of dnaK gene in knockdown strain by qPCR. Values are presented as average plus standard deviation of 3 replicates. For statistical analyses, relative expression values displayed by the knockdown mutant were compared to those presented by the cells containing pMaOri.dCas9 alone (*P < 0.05).
