Genetic elements driving circular RNA translation and methods of use
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsHoward Y. Chang, Chun-Kan Chen
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Abstract
Provided herein are recombinant circular RNA (circRNA) molecules comprising an internal ribosome entry site (IRES) operably linked to a protein-coding nucleic acid sequence. The IRES includes at least one RNA secondary structure element; and a sequence region that is complementary to an 18S ribosomal RNA (rRNA). Methods of producing a protein in a cell using the recombinant circRNA molecules are also provided.
Claims16
A method of inhibiting translation of a protein-coding nucleic acid sequence present on recombinant circular RNA molecule comprising a protein-coding nucleic acid sequence and an internal ribosome entry site (IRES) sequence region operably linked to the protein-coding nucleic acid sequence, wherein the IRES sequence region comprises:
- (i) at least one RNA secondary structure element;
- (ii) a sequence that is complementary to an 18S ribosomal RNA (rRNA);
- and
- wherein the IRES sequence region has a minimum free energy (MFE) of less than −18.9 kJ/mol and a melting temperature of at least 35.0° C.,
- which method comprises contacting the recombinant circular RNA molecule with a first oligonucleotide molecule, whereby said first oligonucleotide molecule hybridizes to the at least one RNA secondary structure element and/or the nucleic acid sequence complementary to an 18S rRNA present in the IRES sequence region of the recombinant circular RNA molecule and inhibits translation of the protein-coding nucleic acid sequence.
Dependencies are inferred from the claim text.