CPC C12N 15/113 (2013.01) [C12N 2310/11 (2013.01); C12N 2310/124 (2013.01); C12N 2310/532 (2013.01)] | 27 Claims |
1. A self-circularization RNA construct comprising, from the 5′-terminus to the 3′-terminus:
a) an internal guide sequence (IGS) region comprising guanine (G) at the 5′-end;
b) a ribozyme region wherein the ribozyme region is a Group I intron ribozyme;
c) a gene of interest (GOI) region; and
d) a fragment containing target splice site comprising uracil (U) at the 3′-end,
wherein the 5′-end G of the IGS region and the 3′-end U of the target splice site-containing fragment form a G : U wobble base pair,
wherein the a) IGS region comprises a sequence of 2 to 10 contiguous nucleotides, which is reverse complementary to a 3′-end portion nucleotide sequence of the d) target splice site-containing fragment, and
wherein the self-circularization RNA construct allows production of a circular RNA comprising the c) gene of interest region and the d) target splice site-containing fragment, but not the b) ribozyme region and the a) IGS region.
|