Controlling Antibiotic Resistant or Highly Virulent Pathogens Through Plasmid Curing
Group II introns are a novel class of catalytic RNA molecule that are capable of site specific insertion into DNA targets. This property can be reengineered to facilitate site specific DNA integration into a wide range of DNA targets. We intend to exploit this property of group II introns to effect plasmid curing by disrupting critical sequences or proteins responsible for plasmid replication/spread or stable maintenance. In a complimentary approach integration of a group II intron into a plasmid will be used to activate a toxic payload that can systematically sense and destroy plasmid bearing cells. If successful in the initial proposed work, during a phase I option we propose to modify the vector backbones on which the group II introns are delivered to a conjugation based system. Conjugative systems can be quite efficient and in some cases approach near 100% efficiency of plasmid transfer. We will port our expression cassettes to plasmids that can be delivered by live organisms that are permitted to be used as probiotics or have gained FDA approval in an IND.
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