System and method for cell type-specific translation of RNA molecules in eukaryotes
AssigneeSrtd Biotech GmbH
InventorsHeribert Bohlen, Bernd Hoffmann
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Abstract
The present technology comprises methods and RNA constructs for the targeted translation of a polypeptide of interest in a eukaryotic target cell, and to the use thereof in therapeutic clinical applications.
Claims14
Method for the targeted translation of a polypeptide of interest in a eukaryotic target cell, characterized by introducing an RNA construct into cells, the RNA construct having at least the following elements in the 5′ to 3′ direction:
- (a) an antisense sequence (a) that is complementary to at least a portion of an mRNA of a gene expressed by the target cell;
- (b) a blocking element (b) that interacts with at least a portion of the translation initiator element (d);
- (c) a stabilizing element (c), wherein the stabilizing element is a nucleotide sequence that prevents degradation of RNA in the 3′ direction from the stabilizing element (c) by exonucleases, the stabilizing element (c) comprising binding partners and/or secondary structures configured to restrict complete degradation of the RNA in the 3′ direction from the stabilizing element (c);
- (d) a translation initiator element (d);
- (e) a sequence (e) coding for the polypeptide of interest,
- translation of the polypeptide of interest being prevented in a non-target eukaryotic cell by base pairing of the blocking element (b) with the translation initiator element (d), and translation of the encoded polypeptide taking place in a eukaryotic target cell due to processing of the RNA construct induced by interaction of the antisense sequence with the mRNA expressed by the target cell.
An RNA construct for the targeted translation of a polypeptide of interest in a eukaryotic target cell, wherein the RNA construct has at least the following elements in the 5′ to 3′ direction:
- (a) an antisense sequence (a) that is complementary to at least a portion of an mRNA of a gene expressed by the target cell;
- (b) a blocking element (b) that is complementary to or otherwise inhibits at least a portion of the translation initiator element (d);
- (c) a stabilizing element (c) wherein the stabilizing element (c) is a nucleotide sequence that prevents degradation of RNA in the 3′ direction from the stabilizing element (c) by exonucleases, the stabilizing element (c) comprising binding partners and/or secondary structures configured to restrict complete degradation of the RNA in the 3′ direction from the stabilizing element (c);
- (d) a translation initiator element (d) that functions independently of 5′ cap and is active only in target cells;
- (e) a sequence (e) coding for the polypeptide of interest;
- wherein the RNA construct is configured such that translation of the polypeptide of interest is prevented in a non-target eukaryotic cell when the RNA construct is in said non-target eukaryotic cell by base pairing of the blocking element (b) with the translation initiator element (d), and translation of the encoded polypeptide in a eukaryotic target cell takes place when the RNA construct is in said eukaryotic target cell due to processing of the RNA construct induced by interaction of the antisense sequence (a) with the mRNA expressed by the target cell as a result of which the translation initiator element is activated.
Dependencies are inferred from the claim text.