Methods and compositions for needleless delivery of macromolecules

A polynucleotide that encodes a delivery construct, said delivery construct comprising:

• a)—a receptor binding domain,
• b)—a transcytosis domain,
• c)—a macromolecule to be delivered to a subject, and
• d)—a cleavable linker,
• wherein cleavage at said cleavable linker separates said macromolecule from the remainder of said construct, and wherein said cleavable linker is cleavable by an enzyme that i) exhibits greater activity at a basal-lateral membrane of a polarized epithelial cell of said subject than at an apical membrane of the polarized epithelial cell, or ii) exhibits greater activity in the plasma of said subject than at an apical membrane of the polarized epithelial cell of the subject.

A polynucleotide that encodes a delivery construct, said polynucleotide comprising:

• a)—a nucleic acid sequence encoding a receptor binding domain,
• b)—a nucleic acid sequence encoding a transcytosis domain,
• c)—a nucleic acid sequence encoding a cleavable linker, and
• d)—a nucleic acid sequence comprising a polylinker insertion site,
• wherein said polylinker insertion site is oriented relative to said nucleic acid sequence encoding a cleavable linker to allow to cleavage of the cleavable linker to separate a macromolecule that is encoded by a nucleic acid inserted into said polylinker insertion site from the remainder of said delivery construct, and wherein said cleavable linker is cleavable by an enzyme that i) exhibits greater activity at a basal-lateral membrane of a polarized epithelial cell of said subject than at an apical membrane of the polarized epithelial cell, or ii) exhibits greater activity in the plasma of said subject than at an apical membrane of the polarized epithelial cell of the subject.

A composition comprising a delivery construct, said delivery construct comprising:

• a)—a receptor binding domain,
• b)—a transcytosis domain,
• c)—a macromolecule to be delivered to a subject, and
• d)—a cleavable linker,
• wherein said cleavable linker is cleavable by an enzyme that i) exhibits greater activity at a basal-lateral membrane of a polarized epithelial cell of said subject than at an apical membrane of the polarized epithelial cell, or ii) exhibits greater activity in the plasma of said subject than at an apical membrane of the polarized epithelial cell of the subject.