Title: CRISPR/Cas9 in mollicutes : from natural system to genome engineering tools

Abstract: CRISPR/Cas systems are widely represented in bacteria and archaea and provide them an adaptative defense mechanism against exogenous DNA or RNA. Because of its wide use as a genome engineering tool, type II CRISPR system from Streptococcus pyogenes has been extensively studied. The core system includes the endonuclease Cas9 and one sgRNA Cas9 on a target site with a recognition specificity depending on the NGG Protospacer Adjacent Motif (PAM). Mollicutes are minimal bacteria that have no cell wall and most of them use an alternative genetic code. Their genomes are AT rich (~70-75%) with a size ranging from 0.58 to 2.2Mbp. Most of them have a parasitic lifestyle and are pathogens of a wide host diversity including humans, other animals and plants. For most mollicutes species, genome engineering currently remains challenging mainly because homologous recombination is poorly efficient. The aim of my thesis was to (1) characterize CRISPR/Cas9 systems of mollicutes, (2) develop a genome engineering tool based on endogenous CRISPR/Cas9 system and, (3) extend the toolbox for some mycoplasma species of veterinary interest for any precise genetics tool was available.

Biography: Thomas Ipoutcha is a postdoctoral scholar in the Department of Civil and Environmental Engineering, at Northwestern University. He previously worked on the Mollicutes team to develop genome engineering tools on minimal bacteria. He completed his Ph.D. at the University of Bordeaux (France) in February 2022. He is now interested in solving important health issues, like drug-resistant Pseudomonas aeruginosa. He is looking to develop strategies to fight this bacteria using phage therapy technology. You can find him on Twitter and check out his publications via ORCID.