INNATE IMMUNE SUPPRESSION ENABLES REPEATED DELIVERY OF LONG RNA MOLECULES
First Claim
1. A method for suppressing the innate immune response of a cell to transfection with a nucleic acid by introducing to the cell an effective amount of an agent that reduces the expression of one or more proteins in the innate immune response pathway selected from the group consisting of TP53, TLR3, TLR7, RARRES3, IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNK, IFNB1, IL6, TICAM1, TICAM2, MAVS, STAT1, STAT2, EIF2AK2, IRF3, TBK1, CDKN1A, CDKN2A, RNASEL, IFNAR1, IFNAR2, OAS1, OAS2, OAS3, OASL, RB1, ISG15, ISG20, IFIT1, IFIT2, IFIT3, and IFIT5, or to a biologically-active fragment, analog or variant thereof.
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Accused Products
Abstract
The present invention relates in part to methods for suppressing the innate immune response of a cell to transfection with an exogenous nucleic acid, to methods for increasing expression of a protein encoded by an exogenous nucleic acid by repeated delivery of the exogenous nucleic acid to a cell, and to methods of changing the phenotype of a cell by differentiating, transdifferentiating or dedifferentiating cells by repeatedly delivering one or more nucleic acids that encode defined proteins. A method is provided for extended transient transfection by repeated delivery of an in vitro-transcribed RNA (“ivT-RNA”) to a cell to achieve a high and sustained level of expression of a protein encoded by an ivT-RNA transcripts.
157 Citations
52 Claims
- 1. A method for suppressing the innate immune response of a cell to transfection with a nucleic acid by introducing to the cell an effective amount of an agent that reduces the expression of one or more proteins in the innate immune response pathway selected from the group consisting of TP53, TLR3, TLR7, RARRES3, IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNK, IFNB1, IL6, TICAM1, TICAM2, MAVS, STAT1, STAT2, EIF2AK2, IRF3, TBK1, CDKN1A, CDKN2A, RNASEL, IFNAR1, IFNAR2, OAS1, OAS2, OAS3, OASL, RB1, ISG15, ISG20, IFIT1, IFIT2, IFIT3, and IFIT5, or to a biologically-active fragment, analog or variant thereof.
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7. A method for transfecting a cell with a nucleic acid molecule, which comprises
a. suppressing the innate immune response of the cell, and b. introducing the nucleic acid into the cell.
- 32. A composition comprising a plurality of different siRNAs, one or more antisense oligonucleotides, or combinations thereof that specifically hybridize to mRNA or DNA molecules encoding two or more proteins in the innate immune response pathway selected from the group comprising TP53, TLR3, TLR7, RARRES3, IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNK, IFNB1, IL6, TICAM1, TICAM2, MAVS, STAT1, STAT2, EIF2AK2, IRF3, TBK1, CDKN1A, CDKN2A, RNASEL, IFNAR1, IFNAR2, OAS1, OAS2, OAS3, OASL, RB1, ISG15, ISG20, IFIT1, IFIT2, IFIT3, IFIT5 or biologically-active fragments, variants or analogs thereof, or to a DNA or RNA molecule encoding any of the enumerated proteins, thereby reducing expression of the two or more proteins.
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36. A DNA template comprising, a transcription promoter operably linked to a construct that produces an in vitro-transcribed RNA comprising,
(i) a stable 5′ - UTR, and
(ii) a strong Kozak consensus sequence linked to a protein coding sequence (CDS). - View Dependent Claims (37, 38, 39, 40, 41, 42, 43, 44)
- UTR, and
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45. An in vitro-transcribed RNA transcript encoding a protein of interest, comprising in operable combination,
(i) a stable 5′ - UTR, and
(ii) a strong Kozak sequence linked to a protein coding sequence (CDS). - View Dependent Claims (46, 47, 48, 49)
- UTR, and
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50. A kit comprising a DNA template having a strong transcription promoter operably linked to a construct that produces an in vitro-transcribed RNA comprising,
(i) a stable 5′ - UTR, and
(ii) a first restriction site capable of joining the 5′
UTR to a Kozak consensus sequence. - View Dependent Claims (51)
- UTR, and
Specification