Chloroplasts engineered to express pharmaceutical proteins in edible plants
AssigneeThe Trustees of the University of Pennsylvania
InventorHenry Daniell
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Abstract
Disclosed herein are compositions for conferring oral tolerance in mammals to autoimmune disorders such as diabetes, as well as vectors and methods for plastid transformation of plants to produce a CTB-Pins protein for oral delivery. The invention also extends to the transformed plants, plant parts, and seeds and progeny thereof. The invention is applicable to plants edible without torrefying. Exemplified herein is the stable transformation of Lactuca sativa such that CTB-Pins is expressed.
Claims31
A plant cell of an edible plant, wherein said plant cell comprises chloroplasts transformed to express a pharmaceutical protein of interest.
An orally-administrable composition comprising a pharmaceutical protein of interest expressed in a Lactuca sativa chloroplast;
- and rubisco.
A sample of pharmaceutical protein bioencapsulated in chloroplasts of an edible plant cell.
A Lactuca sativa plant plastid comprising a plastid genome transformed with a heterologous DNA coding sequence encoding a polypeptide of interest, and integrated into said plastid genome such that said polypeptide of interest is expressed in and present in said plastid.
A Lactuca sativa plant cell homoplasmic with respect to plastids transformed to express a pharmaceutical protein of interest.
A method of retarding the development of or treating diabetes in a subject in need thereof comprising administering to said subject a composition comprising a CTB-Pins polypeptide expressed in a chloroplast in a plant edible without cooking and a plant remnant.
A composition for retarding the development of or treating diabetes comprising a therapeutically effective amount of a CTB-Pins polypeptide and a plant remnant from Lactuca sativa.
A stable plastid transformation and expression vector for transformation of Lactuca sativa plastid, said vector comprises an expression cassette comprising, as operably linked components in the 5′ to the 3′ direction of translation, a promoter operative in said plastid, a selectable marker sequence, a heterologous polynucleotide sequence coding for comprising at least 70% identity to a CTB-Pins protein, transcription termination functional in said plastid, and flanking each side of the expression cassette, flanking DNA sequences which are homologous to a DNA sequence of the target plastid genome, whereby stable integration of the heterologous coding sequence into the plastid genome of the target plant is facilitated through homologous recombination of the flanking sequence with the homologous sequences in the target plastid genome.
A Lactuca sativa plastid genome transformed to contain a CTB-Pins polynucleotide configured so as to express CTB-Pins protein.
A composition for retarding the development of or treating diabetes comprising a therapeutically effective amount of a CTB-Pins polypeptide and rubisco derived from Lactuca sativa.
A method of producing a CTB-Pins containing composition, said method comprising:
- obtaining a stably transformed Lactuca sativa plant which comprises a plastid stably transformed with an expression vector which comprises an expression cassette comprising, as operably linked components in the 5′ to the 3′ direction of translation, a promoter operative in a plastid, a selectable marker sequence, a heterologous polynucleotide sequence coding for comprising at least 70% identity to a CTB-Pins protein, transcription termination functional in said plastid, and flanking each side of the expression cassette, flanking DNA sequences which are homologous to a DNA sequence of the target plastid genome, whereby stable integration of the heterologous coding sequence into the plastid genome of the target Lactuca sativa plant is facilitated through homologous recombination of the flanking sequence with the homologous sequences in the target plastid genome;
- and
- homogenizing material of said stably transformed Lactuca sativa plant to produce homogenized material.
Dependencies are inferred from the claim text.