Expression cassette including intron for high expression of protein of interest and use thereof

Patent metadata

Field	Value
Publication number	US-12534738-B2
Application number	US-202017784461-A
Country	US
Kind code	B2
Filing date	Dec 21, 2020
Priority date	Dec 20, 2019
Publication date	Jan 27, 2026
Grant date	Jan 27, 2026

Abstract

The present invention relates to an expression cassette comprising an intron, for high expression of a protein of interest, and use thereof. More particularly, the present invention relates to an expression cassette comprising an intron sequence, a vector and transformant including same, and a method for preparing a protein of interest using same. The protein-of-interest expression cassette according to the present invention has the effect of inducing high expression of a protein of interest through a single transduction.

First claim

The invention claimed is:

1 . An expression-enhancing intron, comprising:
an expression-enhancing sequence comprising a splicing donor of SEQ ID NO: 1; a branch sequence of any one of SEQ ID NOs: 4 to 6; and a polyY sequence and a splicing acceptor of any one of SEQ ID NOs: 7 to 12.

2 . The expression-enhancing intron of claim 1 , wherein the expression-enhancing intron is any one of SEQ ID NOs: 20 to 23.

3 . An expression cassette for producing a protein of interest comprising the expression-enhancing intron of claim 1 , a promoter, an intron sequence, a polynucleotide encoding the protein of interest, and a polyA sequence.

4 . The expression cassette of claim 3 , wherein one or more expression-enhancing introns exist on a single protein-of-interest expression cassette.

5 . The expression cassette of claim 3 , wherein the expression-enhancing intron is comprised at one or more positions selected from the group consisting of the following positions:
a) between the promoter and the polynucleotide encoding the protein of interest; b) between the polynucleotide encoding the protein of interest and the poly A; and c) inside the polynucleotide encoding the protein of interest.

6 . The expression cassette of claim 3 , wherein the promoter is a CMV promoter.

7 . The expression cassette of claim 3 , wherein the protein of interest is an antibody or a fragment thereof.

8 . The expression cassette of claim 3 , wherein the polyA sequence is SEQ ID NO: 13 or 14.

9 . The expression cassette of claim 3 , wherein the expression cassette has any one or more effects selected from the group consisting of the following effects by regulating intron splicing: an effect of promoting mRNA export out of the nucleus, and an effect of increasing an efficiency of translation of the protein of interest.

10 . A vector containing the expression cassette for producing a protein of interest according to claim 3 .

11 . A transformant transformed with the vector of claim 10 .

12 . The transformant of claim 11 , wherein the transformant is a eukaryotic cell.

13 . A method for producing a protein of interest comprising a step of culturing the transformant of claim 11 .

Assignees

• Celltrion Inc

Inventors

• Kim Man Su
• Kim Min Soo

Classifications

• C12N2830/50

  regulating RNA stability, not being an intron, e.g. poly A signal · CPC title

• C12N2830/42

  being an intron or intervening sequence for splicing and/or stability of RNA · CPC title

• C07K16/22

  against growth factors {; against growth regulators} · CPC title

• C12N15/85 Primary

  for animal cells · CPC title

• C12P21/00

  Preparation of peptides or proteins (single cell protein C12N1/00) · CPC title