Synthetic bacteriophages and bacteriophage compositions
Disclosed herein are novel synthetic bacteriophages and bacteriophage compositions, methods of production thereof, and therapeutic uses thereof.
1. A synthetic bacteriophage comprising mutations in a tail fiber tip protein, wherein the mutations: (i) are engineered in one or more binding loops of the tail fiber tip protein, wherein the binding loops face the basal or apex side of the tail fiber; and (ii) cause the bacteriophage to have a different host range than the corresponding bacteriophage lacking the mutations in the tail fiber tip protein.
2. The synthetic bacteriophage of claim 1 , wherein the mutations are in one or more of loops BC, DE, FG and/or HI.
3. The synthetic bacteriophage of claim 1 , wherein the bacteriophage is a T3 bacteriophage, a T7 bacteriophage, or a bacteriophage having a tail fiber tip protein that has 90% or greater amino acid identity to a T3 bacteriophage gp17 tail fiber tip protein or a T7 bacteriophage gp17 tail fiber tip protein.
4. The synthetic bacteriophage of claim 1 , wherein the tail fiber tip protein is gp17.
5. A pharmaceutical composition comprising the synthetic bacteriophage as claimed in claim 1 , and a pharmaceutically-acceptable carrier.
6. A pharmaceutical composition comprising two or more types of synthetic bacteriophages as claimed in claim 1 , wherein each of the two or more types of bacteriophages has different mutations in the tail fiber tip protein, and a pharmaceutically acceptable carrier.
7. The composition of claim 6 , wherein the two or more types of bacteriophages have different host ranges.
8. A method for treating a bacterial infection comprising
administering to a subject having a bacterial infection in need of treatment the pharmaceutical composition as claimed in claim 5 .
9. A method of producing one or more synthetic bacteriophages comprising mutating a tail fiber protein of a bacteriophage wherein the mutations: (i) are engineered in one or more binding loops of the tail fiber tip protein, wherein the binding loops face the basal or apex side of the tail fiber; and (ii) cause the bacteriophage to have a different host range than the corresponding bacteriophage lacking the mutations in the tail fiber tip protein.
10. The method of claim 9 , wherein the bacteriophage is a T3 bacteriophage, a T7 bacteriophage, or a bacteriophage having a tail fiber tip protein that has 90% or greater amino acid identity to a T3 bacteriophage gp17 tail fiber tip protein or a T7 bacteriophage gp17 tail fiber tip protein.
11. The method of claim 9 , wherein the tail fiber tip protein is gp17.
12. The method of claim 9 comprising replacing one or more codons within at least one binding loop of a tail fiber protein with the degenerate codon NNK to introduce amino acid variability in the at least one loop.
13. The method of claim 12 , wherein the step of replacing one or more codons comprises amplifying a sequence comprising a bacteriophage gp17 gene region to produce an amplification product.
14. The method of claim 13 , further comprising:
circularizing the amplification product;
introducing the circularized amplification product into bacteriophage susceptible bacterial cells;
infecting the bacterial cells with wild-type bacteriophages, wherein said infection facilitates the recombination of the wild-type bacteriophage with the amplification product; and
isolating recombinant synthetic bacteriophages comprising the amplification product.
15. The method of claim 11 , wherein the amino acid variability is introduced in any one or more of the BC, DE, FG, and/or HI loops of the gp17 tail fiber protein.
16. The method of claim 9 , wherein the mutation in the one or more binding loops comprises one or more amino acid substitutions.
17. The method of claim 9 , wherein the mutation in the one or more binding loops shortens or lengthens the one or more binding loops by at least one amino acid.
18. The synthetic bacteriophage of claim 1 , wherein the bacteriophage is a bacteriophage having a tail fiber tip protein that has 90% or greater amino acid identity to the C-terminal 104 amino acids of T3 bacteriophage gp17 tail fiber tip protein or the C-terminal 99 amino acids of T7 gp17 tail fiber tip protein.