Target validation of putative histidine-kinase inhibitors using a CRISPR/dCas9 approach


Recently identified putative histidine kinase inhibitors (Velikova et al. 2016, Sci Rep) loaded to nanoparticles for delivery of antibacterials to Gram negatives (Gr-) are bactericidal against Gr- such as  e.g. E. coli despite the lack of essential histidine kinases (HK) in this pathogen (Velikova et al. 2017, Nanomedicine). Therefore, we hypothesized the observed bactericidal effect could be due to the simultaneous inhibition of multiple targets. To test this hypothesis we have employed a CRISPR/dCas9 approach.


To study the effect of the simultaneous knock down of multiple genes in a Gr- pathogen.

Methods :

Molecular cloning

Molecular microbiology

General microbiology techniques

Aseptic work


Nadya Velikova,