Nucleic acid molecule comprising asymmetrically modified ITR for improving expression rate of inserted gene, and use thereof

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.