# US 11,993,782 B2 · Lentivirus packaging system, lentivirus produced by the same, cell transduced by the lentivirus, method for improving lentivirus production in a host cell, and method of using the cell for treating cancer

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,993,782 B2
- Document ID: US11993782B2
- Status: Granted
- Kind: B2
- Application number: 17/377,830
- Filed: 2021-07-16
- Published: 2022-11-03
- Granted: 2024-05-28
- Earliest claimed: 2021-04-28
- Assignees: Pell Bio-Med Technology Co., Ltd.
- Applicants: Pell Bio-Med Technology Co., Ltd.
- Inventors: Wei-Chi Lin; Ssu-Yu Chou; Yao-Cheng Yang; Chien-Ting Lin; Chen-Lung Lin
- Web page: https://patentgrep.com/patent/US11993782B2

## CPC

- C12N 15/86: Viral vectors
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/4211: {CD19 or B4}
- C07K 14/70503: {Immunoglobulin superfamily}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12N 2740/15043: viral genome or elements thereof as genetic vector
- C12N 2740/16043: viral genome or elements thereof as genetic vector
- C12N 2740/16052: relating to complementing cells and packaging systems for producing virus or viral particles
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

Provided is a lentivirus packaging system, which comprises: a transfer plasmid comprising a nucleotide sequence of TAR-reserved-chimeric 5′ long terminal repeat (LTR); at least one packaging plasmid comprising a nucleotide sequence encoding TAR RNA binding protein, a nucleotide sequence of rev gene, a nucleotide sequence of gag gene, and a nucleotide sequence of pol gene; and an envelope plasmid. Due to the expression of gene of TAR RNA binding protein by the packaging plasmids, the produced lentivirus has higher virus titer and can improve the transduction rate and the gene delivery efficiency during cell transduction. The present invention further provides a method of improving lentivirus production in a host cell, which comprises using the lentivirus packaging system to transfect the host cell. The present invention further provides a cell transduced by the lentivirus and a method of using the cell for treating cancer.

## Claims

### Claim 1

A lentivirus packaging system, comprising:
a transfer plasmid comprising a nucleotide sequence of a trans-activation response (TAR) element-reserved chimeric 5′ long terminal repeat (TAR-reserved chimeric 5′-LTR);
a first packaging plasmid comprising a CMV promoter, a nucleotide sequence of a gag gene and a nucleotide sequence of a pol gene;
a second packaging plasmid comprising a CMV promoter, a nucleotide sequence of a tat gene;
a third packaging plasmid comprising an OG-RSV promoter, a nucleotide sequence of a rev gene; and
an envelope plasmid,
wherein a weight ratio of the transfer plasmid, the first packaging plasmid, the second packaging plasmid, the third packaging plasmid and the envelope plasmid is 3 to 12:3 to 7:1 to 4:1 to 4:0.8 to 6, respectively.

### Claim 2 (depends on 1)

The lentivirus packaging system as claimed in claim 1, wherein the nucleotide sequence of the tat gene comprises the nucleotide sequence of SEQ ID NO: 1.

### Claim 3 (depends on 1)

The lentivirus packaging system as claimed in claim 1, wherein the envelope plasmid comprises a nucleotide sequence encoding a vesicular stomatitis virus glycoprotein (VSV-G) or a nucleotide sequence encoding a baboon endogenous virus envelope.

### Claim 4 (depends on 1)

The lentivirus packaging system as claimed in claim 1, wherein the nucleotide sequence of the TAR-reserved chimeric 5′-LTR has a nucleotide sequence of a TAR, and the nucleotide sequence of the TAR comprises the nucleotide sequence of SEQ ID NO: 2.

### Claim 5 (depends on 1)

A method of increasing the yield of lentivirus in a host cell, comprising a step of using the lentivirus packaging system as claimed in claim 1 to transfect the host cell, wherein the host cell is a pkr knockout 293T cell.

### Claim 6 (depends on 1)

A method of increasing the yield of lentivirus in a host cell, comprising a step of using the lentivirus packaging system as claimed in claim 1 to transfect the host cell.

### Claim 7 (depends on 6)

The method of increasing the yield of lentivirus in the host cell as claimed in claim 6, wherein the host cell comprises a mammalian cell.

### Claim 8 (depends on 7)

The method of increasing the yield of lentivirus in the host cell as claimed in claim 7, wherein the mammalian cell comprises a Human Embryonic Kidney 293 cell (HEK293) or a 293T cell (HEK293T).

### Claim 9 (depends on 1)

An isolated cell transfected with the lentivirus packaging system as claimed in claim 1.