
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.
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R106
Paving the Road Towards Universal Microbial Genome Editing
Constantinos Patinios, PhD (VU-EMBL Partnership Institute for Genome Editing Technologies)
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