Modifying bacteriophage
Inventors
Fairhead, Heather • Wilkinson, Adam • BARNARD, Anne • SEVERI, Emmanuele • Anderson, Neil • Pitts, Katy
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-10953052-B2
Publication Date
2021-03-23
Expiration Date
Abstract
A method for modifying the genome of a lytic target phage, uses of the method and products thereof are described. Compositions comprising such phage are also described. The compositions may be formulated as a medicament, which are useful for human treatment and may treat various conditions, including bacterial infections.
Core Innovation
The invention provides a method for modifying the genome of a lytic target phage whose genome includes a first target sequence and a second target sequence. The method uses a vector containing a phage-targeting region comprising a host range determinant of a marker phage that is different from the host range determinant of the target phage, where the phage-targeting region is flanked by first and second flanking sequences homologous to the first and second target sequences of the target phage genome.
A vector is introduced into a first host cell that is a host for the target phage, and the first host cell is infected with the target phage. The method allows replication of the phage and recombination between the target phage genome and the vector, so that the genome of the target phage is modified to incorporate the exogenous DNA sequence.
Resultant phage are then propagated on a second host cell that is a host for the marker phage and not the target phage. Propagation on the second host cell indicates that the resultant phage comprise the exogenous DNA sequence, after which the resultant phage are harvested.
Claims Coverage
The independent claim defines the full method for modifying the genome of a lytic target phage using a marker-phage host range determinant selection strategy. The dependent claims refine specific structural and sequence aspects of the host range determinant region and the exogenous DNA payload, and add additional genome-modification constraints.
Marker phage host range selection with homologous flanking phage-targeting region
A method for modifying the genome of a lytic target phage using a vector whose phage-targeting region comprises a host range determinant of a marker phage, different from the host range determinant of the target phage, wherein the phage-targeting region is flanked by first and second flanking sequences homologous to the first and second target sequences of the target phage genome.
Exogenous DNA incorporation via recombination during phage replication
The vector further comprises an exogenous DNA sequence for incorporation into the genome of the target phage, and replication of the phage and recombination between the target phage genome and vector occurs to modify the genome of the target phage.
Two-host propagation that selects for incorporation by marker host range determinant
Resultant phage are propagated on a second host cell that is a host for the marker phage and not the target phage, wherein the propagation indicates that the resultant phage comprise the exogenous DNA sequence, followed by harvesting the resultant phage.
Overall claim coverage centers on a recombination-based genome modification of a lytic target phage using a vector with homologous flanking sequences and a marker phage host range determinant different from the target phage, coupled with propagation on a second host that supports the marker but not the target to indicate incorporation of the exogenous DNA.
Stated Advantages
Documented Applications
No documented applications found
Interested in licensing this patent?