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US 11,351,271 B2Granted
RNA-based logic circuits with RNA binding proteins, aptamers and small molecules
AssigneesMassachusetts Institute of Technology, Kyoto University
InventorsRon Weiss, Liliana Wroblewska, Velia Siciliano, Tasuku Kitada, Maria Hottelet Foley, Katie Bodner, Hirohide Saito, Kei Endo, Darrell J. Irvine, Tyler Wagner, Jacob Becraft
Life of this patent
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Abstract
Engineered synthetic RNA-based genetic circuits are provided that are regulated exclusively at the post-transcriptional level.
Claims15
A synthetic RNA circuit comprising
- (a) a first RNA molecule comprising a 5′ cap, at least one target site specific for a first microRNA molecule, a sequence encoding an RNA binding protein, and a poly(A) tail;
- and
- (b) a second RNA molecule comprising a 5′ cap, at least one target site specific for a second microRNA molecule, at least one RNA motif that is capable of being bound by the RNA binding protein in (a), a sequence encoding an output molecule, and a poly(A) tail,
- wherein the first microRNA molecule and the second microRNA molecule are different, wherein the first microRNA molecule is a microRNA that is present in a first cell type such that it reduces expression of the first RNA molecule in the first cell type and the second microRNA molecule is a microRNA that is present in a second cell type such that it reduces expression of the second RNA molecule in the second cell type but does not reduce expression of the second RNA molecule in the first cell type or reduces expression of the second RNA molecule less in the first cell type compared to the second cell type, wherein the first cell type and the second cell type are different, wherein the RNA binding protein inhibits production of the output molecule when bound to the at least one RNA motif, and wherein the synthetic RNA circuit is capable of being expressed in a eukaryotic cell.
1 more element
A synthetic RNA circuit comprising:
- (a) a first RNA molecule comprising a 5′ cap, at least one first RNA motif, a sequence encoding a second RNA binding protein that is capable of binding to a second RNA motif and that is capable of inhibiting protein production, at least one target site specific for a first siRNA molecule or first microRNA molecule, and a poly(A) tail;
- and
- (b) a second RNA molecule comprising a 5′ cap, at least one second RNA motif that is capable of being bound by the second RNA binding protein, a sequence encoding a first RNA binding protein that is capable of binding to the first RNA motif and inhibiting protein production, at least one target site specific for a second siRNA molecule or second microRNA molecule, and a poly(A) tail, wherein the first siRNA molecule or first microRNA molecule and the second siRNA molecule or second microRNA molecule are different, and wherein the first RNA motif and the second RNA motif are different wherein the first RNA binding protein, upon binding to the first RNA motif, inhibits expression of the second RNA binding protein, and wherein the second RNA binding protein, upon binding to the second RNA motif, inhibits expression of the first RNA binding protein, and wherein the synthetic RNA circuit is capable of being expressed in a eukaryotic cell.
Dependencies are inferred from the claim text.