Method of producing transduction particles

Inventors

SEMSEY, Szabolcs

Abstract

The invention relates to the production of transduction particles, such as phage, as well as compositions comprising the particles and use of these. The particles are particularly useful for delivering toxic payloads into target bacteria for antibacterial action. The invention is also useful for the production of wild-type (“native”) phage and phage cocktails comprising different types of phage. Embodiments enable production of compositions of such particles for use in medical, environmental or food production settings.

Core Innovation

The invention relates to producing transduction particles that recognize a receptor on bacterial target cells for transduction of the cells. The particles are produced in bacterial producer cells that do not express the receptor on their surface, and this production configuration enhances the production yield compared to producing the transduction particles in bacterial producer cells that express the receptor on their surface.

The described transduction-particle embodiments include phage-based transduction particles and non-self-replicative transduction particle systems. In these embodiments, the documents describe the use of phage capsids and packaging signals, and the inclusion of a nucleic acid sequence of interest encoding antibacterial payloads, including CRISPR/Cas systems with guide RNA and a Cas nuclease.

The disclosure also describes addressing receptor features on bacterial cell surfaces, with particular emphasis on receptors associated with cell-surface structures such as LPS. An experimental Example is described in which producer-cell receptor binding is reduced by knocking out an LPS core biosynthesis gene, resulting in a substantially increased transduction particle yield compared to receptor-expressing producer cells.

Claims Coverage

The partial content identifies one independent claim directed to a method of producing transduction particles with enhanced production yield by preventing receptor surface expression in bacterial producer cells. One inventive feature is extracted from that independent claim.

Across the provided claim material, the core claim coverage focuses on producing receptor-recognizing transduction particles in producer cells that lack surface expression of the receptor to increase production yield. Dependent refinements are noted in the partial content, but the main inventive feature extracted here is the receptor non-expression in producer cells paired with enhanced yield.

Stated Advantages

Documented Applications