	US 11,993,783 B1
	Nucleic acid molecule comprising asymmetrically modified ITR for improving expression rate of inserted gene, and use thereof
Suk Chul Bae, Cheongju-si (KR); You Soub Lee, Cheongju-si (KR); Xinzi Chi, Cheongju-si (KR); Seo Yeong Yoo, Cheongju-si (KR); and Woo-Jin Kim, Seoul (KR)
Assigned to GENECRAFT INC., Cheongju-si (KR)
Filed by GENECRAFT INC., Cheongju-si (KR)
Filed on Mar. 27, 2023, as Appl. No. 18/190,590.
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/864 (2006.01); A61K 48/00 (2006.01); C12N 15/113 (2010.01); C12N 15/86 (2006.01); C12N 15/90 (2006.01)

CPC C12N 15/86 (2013.01) [C12N 15/90 (2013.01); C12N 2750/14122 (2013.01); C12N 2750/14143 (2013.01)]

11 Claims

1. A nucleic acid molecule comprising:

a gene expression cassette between a first inverted terminal repeat (ITR) and a second ITR,

wherein the gene expression cassette comprises a heterologous polynucleotide sequence,

wherein:

the first ITR is an adeno-associated virus (AAV) wild-type TTR, and

the second ITR consists essentially of a nucleotide sequence or a complementary sequence thereof, the nucleotide sequence of the second ITR having at least about 90% sequence identity with a nucleotide sequence of any one of SEQ ID NOs: 1 to 9,

wherein the second ITR comprises a terminal resolution site (trs) sequence and a rep-binding element (RBE) sequence,

wherein in the second ITR, all of B, B′, C, C′, and RBE′ regions are deleted, and either A or A′ region, whichever is closer to 3′-end of the (+) strand of the transgene or 5′-end of the (−) strand of the transgene, is deleted,

wherein the second ITR exists as an open-end without forming any of the stem region and the loop region of the stem-loop structure, and

wherein the second ITR retains functional activities of the trs and the RBE.