Targeted rna editing by leveraging endogenous adar using engineered rnas
AssigneePeking University
InventorsWensheng WEI, Zongyi YI, Liang QU, Feng TIAN, Chunhui WANG, Shiyou ZHU, Zhuo ZHOU
Life of this patent
Estimating the term…
Abstract
Provided are methods for editing RNA by introducing a deaminase-recruiting RNA in a host cell for deamination of an adenosine in a target RNA, deaminase-recruiting RNAs used in the RNA editing methods, compositions and kits comprising the same.
Claims34
A method for editing a target RNA in a host cell, comprising introducing a deaminase-recruiting RNA (dRNA) or a construct comprising a nucleic acid encoding the dRNA into the host cell, wherein:
- (1) the dRNA comprises a targeting RNA sequence that is at least partially complementary to the target RNA,
- (2) the dRNA is capable of recruiting an adenosine deaminase acting on RNA (ADAR), and
- (3) the dRNA is a circular RNA or capable of forming a circular RNA.
2-7. (canceled)
A method for editing a target RNA in a host cell, comprising introducing a deaminase-recruiting RNA (dRNA) or a construct comprising a nucleic acid encoding the dRNA into the host cell, wherein the dRNA comprises:
- (1) a targeting RNA sequence that is at least partially complementary to the target RNA and
- (2) a small nucleolar RNA (snoRNA) sequence linked to the 3′ and/or 5′ ends of the targeting RNA sequence;
- and wherein the dRNA is capable of recruiting an adenosine deaminase acting on RNA (ADAR).
A method for editing a target RNA in a host cell, comprising introducing a construct comprising a nucleic acid encoding a deaminase-recruiting RNA (dRNA) into the host cell, wherein:
- (1) the dRNA comprises a targeting RNA sequence that is at least partially complementary to the target RNA,
- (2) the dRNA is capable of recruiting an adenosine deaminase acting on RNA (ADAR), and
- (3) the construct comprises a polymerase II promoter (“Pol II promoter”) operably linked to the nucleic acid encoding the dRNA.
(canceled)
(canceled)
26-32. (canceled)
A construct, comprising a nucleic acid encoding a dRNA for editing a target RNA comprising a targeting RNA sequence that is at least partially complementary to the target RNA, wherein the dRNA is capable of recruiting an Adenosine Deaminase Acting on RNA (ADAR), and wherein the dRNA is circular or is capable of forming a circular RNA, and wherein the construct further comprises a 3′ twister ribozyme sequence linked to the 3′ end of the nucleic acid encoding the dRNA and a 5′ twister ribozyme sequence linked to the 5′ end of the nucleic acid encoding the dRNA, optionally wherein:
- (i) the 3′ twister sequence is twister P3 U2A and the 5′ twister sequence is twister P1;
- or
- (ii) the 5′ twister sequence is twister P3 U2A and the 3′ twister sequence is twister P1.
A deaminase-recruiting RNA (dRNA) for editing a target RNA comprising:
- (1) a targeting RNA sequence that is at least partially complementary to the target RNA and
- (2) a small nucleolar RNA (snoRNA) sequence at the 3′ and/or 5′ ends of the targeting RNA sequence;
- wherein the dRNA is capable of recruiting an adenosine deaminase acting on RNA (ADAR).
(canceled)
38-41. (canceled)
Dependencies are inferred from the claim text.