Functionalization of endogenous bacteria

Abstract

Various aspects and embodiments of the present disclosure are directed to methods and compositions for functionalizing endogenous bacteria in vivo. The methods include delivering to endogenous bacterial cells a recombinant bacteriophage or phagemid that is engineered to contain at least one genetic circuit.

1 .- 21 . (canceled) 
     
     
         22 . A recombinant bacteriophage that is engineered for in vivo delivery to endogenous bacterial cells to contain at least one genetic circuit, wherein the at least one genetic circuit does not express an antimicrobial protein. 
     
     
         23 . The recombinant bacteriophage of  claim 22 , wherein the recombinant bacteriophage is a non-lytic recombinant bacteriophage. 
     
     
         24 . The recombinant bacteriophage of  claim 23 , wherein the non-lytic recombinant bacteriophage is of a family selected from Myoviridae, Siphoviridae, Podoviridae, Tectiviridae, Corticoviridae, Lipothrixviridae, Plasmaviridae, Rudiviridae, Fuselloviridae, Inoviridae, Microviridae, Leviviridae and Cystoviridae. 
     
     
         25 . The recombinant bacteriophage of  claim 24 , wherein the non-lytic recombinant bacteriophage is an Inoviridae M13 or M13-like bacteriophage. 
     
     
         26 . The recombinant bacteriophage of  claim 22 , wherein the at least one genetic circuit contains a nucleic acid with a promoter operably linked to a nucleotide sequence encoding a gene product. 
     
     
         27 . The recombinant bacteriophage of  claim 26 , wherein the nucleic acid is a recombinant nucleic acid. 
     
     
         28 . The recombinant bacteriophage of  claim 26 , wherein the nucleic acid is a synthetic nucleic acid. 
     
     
         29 . The recombinant bacteriophage of  claim 26 , wherein the promoter is a constitutively active promoter. 
     
     
         30 . The recombinant bacteriophage of  claim 26 , wherein the promoter is an inducible promoter. 
     
     
         31 . The recombinant bacteriophage of  claim 26 , wherein the at least one genetic circuit is a recombinase-based genetic circuit. 
     
     
         32 . The recombinant bacteriophage of  claim 22 , wherein the at least one genetic circuit is engineered to express a therapeutic molecule. 
     
     
         33 . The recombinant bacteriophage of  claim 32 , wherein the at least one genetic circuit comprises a nucleic acid with a promoter operably linked to a nucleotide sequence that encodes the therapeutic molecule. 
     
     
         34 . The recombinant bacteriophage of  claim 32 , wherein the therapeutic molecule is an antibody, antibody-based drug, Fc fusion protein, anticoagulant, blood factor, bone morphogenetic protein, engineered protein scaffold, enzyme, growth factor, hormone, interferon, interleukin or thrombolytic. 
     
     
         35 . The recombinant bacteriophage of  claim 22 , wherein the at least one genetic circuit is engineered to detect a condition. 
     
     
         36 . The recombinant bacteriophage of  claim 35 , wherein the at least one genetic circuit comprises a nucleic acid with an inducible promoter operably linked to a nucleotide sequence that encodes a reporter molecule. 
     
     
         37 . The recombinant bacteriophage of  claim 35 , wherein the condition is a cancer, an immune disorder or an infection. 
     
     
         38 . The recombinant bacteriophage of  claim 36 , wherein the reporter molecule is fluorescent protein. 
     
     
         39 . The recombinant bacteriophage of  claim 22 , wherein the at least one genetic circuit comprises a nucleic acid with a promoter operably linked to a nucleotide sequence that encodes toluene dioxygenase or styrene monoxygenase. 
     
     
         40 . A composition comprising the recombinant bacteriophage of  claim 22 . 
     
     
         41 .- 76 . (canceled)