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Mariana Sanchez Londoño

Doctoral Researcher, WG Randau
Biology, Philipps-Universität Marburg

Project: Live-cell single-molecule visualization of the Type IV-A CRISPR-Cas interference mechanism

Project summary:
My project focuses on the activity of Type IV CRISPR-Cas systems, one specialized version of RNA-guided adaptive immunity in bacteria and archaea. Our model organism is Pseudomonas oleovorans which harbours a unique Type IV-A CRISPR-Cas system on its megaplasmid. It exhibits a preference for plasmid targeting and also contains a CRISPR RNA that enables recognition of its own chromosome. Unlike typical CRISPR-Cas systems, target recognition results in the downregulation of target gene expression without DNA degradation. Although recent structural analyses of the effector complex have provided valuable insights, the precise interference mechanism is not fully understood. Therefore, In the wrap-up phase of my project, we aim to elucidate Type IV CRISPR-Cas-mediated plasmid interference using single-molecule microscopy. In collaboration with the research groups of Peter Graumann and Martin Thanbichler, we employ single molecule tracking strategies to visualize the interference process in vivo. These studies are expected to offer insights into the interplay of CRISPR-Cas activity with general cellular processes, aiding CRISPRi tool development to control transcription and plasmid replication, relevant against multi-drug resistance spread.

Cooperation with: Peter Graumann (UMR), Martin Thanbichler (UMR)

Cooperation Partners