Regulation of gene expression by aptamer-modulated RNase p cleavage
AssigneeMeiragtx UK II Limited
InventorXuecui Guo
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Abstract
The present disclosure provides polynucleotide constructs for the modulation of target gene expression by aptamer-mediated ribonuclease cleavage of the target gene RNA and methods of using the constructs to modulate gene expression in response to the presence or absence of a ligand that binds the aptamer. The polynucleotide constructs contains a ribonuclease substrate sequence (e.g., an RNase P substrate) and a riboswitch comprising an effector region and an aptamer such that when the aptamer binds a ligand, target gene expression occurs.
Claims25
A polynucleotide cassette for the regulation of the expression of a target gene the cassette encoding an RNase P substrate sequence linked to a riboswitch, wherein:
- the RNase P substrate sequence comprises a sequence encoding a tRNA, mascRNA, MEN beta tRNA-like structure, or a viral tRNA-like structure—that can initiate RNase P cleavage;
- wherein the RNase P substrate sequence comprises a leader sequence and an acceptor stem;
- the riboswitch is recombinant and comprises an effector region linked to an aptamer sequence;
- the aptamer can specifically bind a small molecule ligand;
- the aptamer undergoes a conformational change when it binds the small molecule ligand;
- the effector region comprises sequence complementary to a portion of the RNase P substrate sequence, the portion comprising leader sequence and/or acceptor stem sequence;
- and
- the effector region sequence is linked to the aptamer such that the conformational change in the aptamer results in formation of a stem comprising the RNase P substrate sequence and the portion of the effector region complementary thereto, wherein the stem can inhibit cleavage of the substrate by RNase P.
Dependencies are inferred from the claim text.