Method for introducing and expressing RNA in animal cells
AssigneeUniversity of Maryland Biotechnology Institute
InventorsDavid M. Hone, George Lewis, Robert Powell
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Abstract
The invention pertains to a method for introducing RNA molecules into eukaryotic cells, wherein the RNA molecules are capable of being translated in the eukaryotic cells or is an antisense RNA or a catalytic RNA, as well as to such bacteria, compositions comprising such bacteria, and nucleic acids which can be introduced into bacteria for practicing the method of the invention. Examples of products the RNA molecule may encode include vaccine antigens, therapeutic agents, immunoregulatory agents or anti-sense RNA molecules or catalytic RNA molecules.
Claims50
A live-invasive bacterium, wherein said bacterium comprises a DNA molecule encoding a therapeutic protein operably linked to a first prokaryotic promoter or bacteriophage promoter such that an RNA molecule encoding said therapeutic protein is transcribed in the bacterium, wherein said RNA molecule comprises a eukaryotic translation element such that said RNA molecule is translated in a eukaryotic host cell, but is not translated in the bacterium, wherein said eukaryotic translation element is a Cap Independent Translation Enhancer (CITE) sequence.
A method for introducing and expressing an RNA molecule encoding a therapeutic protein in animal cells comprising infecting said animal cells with live-invasive bacteria, wherein said bacteria comprise a DNA molecule encoding a therapeutic protein operably linked to a first prokaryotic promoter or bacteriophage promoter such that an RNA molecule encoding said therapeutic protein is transcribed from said DNA molecule in the bacteria, wherein said RNA molecule comprises a eukaryotic translation element such that said RNA molecule is translated in said animal cells, but is not translated in the bacteria, wherein said eukaryotic translation element is a Cap Independent Translation Enhancer (CITE) sequence, and wherein said animal cells are cultured in vitro.
A method for immunizing a subject comprising infecting said subject with attenuated live-invasive bacteria, wherein said bacteria comprise a DNA molecule encoding a vaccine antigen operably linked to a first prokaryotic promoter or bacteriophage promoter such that an RNA molecule encoding said vaccine antigen is transcribed in the bacteria, wherein said RNA molecule comprises a eukaryotic translation element such that said RNA molecule is translated in said subject, but is not translated in the bacteria, wherein said eukaryotic translation element is a Cap Independent Translation Enhancer (CITE) sequence, and wherein said vaccine antigen is expressed at levels sufficient to induce an immune response in the subject.
Dependencies are inferred from the claim text.