VU-EMBL Partnership Institute for Genome Editing Technologies

Constantinos Patinios, PhD

Group Leader
Topic: Paving the Road Towards Universal Microbial Genome Editing Microbe genome editing remains constrained by complex, time-consuming, and often inefficient protocols. These attributes render genome editing, particularly in non-model organisms, a technically challenging and frequently unsuccessful endeavour. In my group at VU-EMBL PI for genome editing technologies, we reconstruct the genome editing pipeline by developing simple, efficient and widely applicable genome editing tools. As an example, I will showcase SIBR-Cas and Append Editing. SIBR is a gene expression system which can control the expression of any gene of interest in virtually any mesophilic prokaryote of interest. We used SIBR to control the expression of the CRISPR-Cas9 and CRISPR-Cas12a proteins in various wild type bacteria, achieving high editing efficiencies. Append Editing is a novel CRISPR-based tool that appends bulky ADP-ribosyl moieties to DNA bases, causing stall of DNA replication and resulting in DNA recombination. We used Append Editing to install small size (~90 bp) substitutions, insertions or deletions in the genome of E. coli, with high editing efficiency and without a reduction in resulting colonies. Moreover, we tested Append Editing in eukaryotic cells (yeast, tobacco and human cells) and noticed that, in contrast to bacteria, site-specific ADP-ribosylation drives replacement of the modified DNA base.

    All Sessions by: Constantinos Patinios, PhD

    • 09:50-10:30 R106

      Paving the Road Towards Universal Microbial Genome Editing

      Constantinos Patinios, PhD (VU-EMBL Partnership Institute for Genome Editing Technologies)