	US 11,685,924 B2
	Genetic elements driving circular RNA translation and methods of use
Howard Y. Chang, Stanford, CA (US); and Chun-Kan Chen, Stanford, CA (US)
Assigned to The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
Filed by The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
Filed on Oct. 3, 2022, as Appl. No. 17/937,617.
Application 17/937,617 is a division of application No. 17/696,606, filed on Mar. 16, 2022, granted, now 11,560,567.
Application 17/696,606 is a continuation of application No. PCT/US2021/039127, filed on Jun. 25, 2021.
Claims priority of provisional application 63/186,507, filed on May 10, 2021.
Claims priority of provisional application 63/043,964, filed on Jun. 25, 2020.
Prior Publication US 2023/0125935 A1, Apr. 27, 2023
Int. Cl. C07H 21/02 (2006.01); C07H 21/04 (2006.01); C12N 15/67 (2006.01); C12N 15/113 (2010.01); C12N 15/85 (2006.01)

CPC C12N 15/67 (2013.01) [C12N 15/113 (2013.01); C12N 15/85 (2013.01); C12N 2840/203 (2013.01)]

16 Claims

1. 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.