Methods to increase transgene expression from bacterial-based delivery systems by co-expressing suppressors of the eukaryotic type I interferon response
Bacterial delivery systems with improved transgene expression are provided. The recombinant bacterial delivery systems deliver transgenes of interest and suppressors of the eukaryotic Type I interferon response to eukaryotic cells. Suppression of the eukaryotic Type I interferon response allows improved expression of the encoded transgene.
1. A method of increasing the production of one or more gene products of interest in a cell or tissue, comprising the step of
administering to said cell or tissue a genetically engineered bacterium comprising nucleic acid sequences encoding: i) said one or more gene products of interest and ii) one or more factors that inhibit a mammalian type I interferon response,
wherein said nucleic acid sequences encoding said one or more gene products of interest are operably linked to a eukaryotic promoter, and said nucleic acid sequences encoding said one or more factors that inhibit a mammalian type I interferon response are operably linked to a eukaryotic promoter or a prokaryotic promoter,
and wherein said step of administering is carried out under conditions which allow said genetically engineered bacterium to invade said cell or tissue, and which allow said cell or tissue to produce said one or more gene products of interest.
2. The method of claim 1 , wherein said nucleic acid sequences encoding said one or more factors that inhibit a mammalian type I interferon response are operably linked to a eukaryotic promoter.
3. The method of claim 1 , wherein said nucleic acid sequences encoding said one or more factors that inhibit a mammalian type I interferon response are operably linked to a prokaryotic promoter.
4. The method of claim 1 , wherein said nucleic acid sequences encoding said one or more factors that inhibit a mammalian type I interferon response are present on a chromosome of said genetically engineered bacterium.
5. The method of claim 1 , wherein one or both of:
i) said nucleic acid sequences encoding said one or more gene products of interest, and
ii) said nucleic acid sequences encoding said one or more factors that inhibit a mammalian type I interferon response,
are present on a plasmid.
6. The method of claim 1 , wherein said one or more factors that inhibit a mammalian type I interferon response are of viral origin.
7. The method of claim 1 , wherein said one or more gene products of interest are Mycobacterium tuberculosis antigens.
8. The method of claim 1 , wherein said genetically engineered bacterium is a bacterium selected from the group consisting of Shigella, Listeria, Salmonella , and Bacille Calmette-Guerin (BCG).
9. The method of claim 1 , wherein said one or more passenger genes is a heterologous transgene.