# US 2026/0092284 A1 · Adeno-associated virus (aav) producer cell lines

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

- Number: US 2026/0092284 A1
- Document ID: US20260092284A1
- Status: Published application
- Kind: A1
- Application number: 19/300,921
- Filed: 2025-08-15
- Published: 2026-04-02
- Earliest claimed: 2023-10-31
- Assignees: Lonza Houston, Inc.
- Applicants: Lonza Houston, Inc.
- Inventors: Bingnan GU; Gang LI; Caitlin TRIPP
- Web page: https://patentgrep.com/patent/US20260092284A1

## CPC

- C12N 15/635: {Externally inducible repressor mediated regulation of gene expression, e.g. tetR inducible by tetracyline}
- C07K 14/005: from viruses
- C12N 15/85: for animal cells
- C12N 15/86: Viral vectors
- C12N 2710/16222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2760/14122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2830/003: tet inducible
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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

The present disclosure relates to nucleic acids encoding helper genes and adeno-associated virus (AAV) genes, under the control of inducible promoters. The disclosure also relates to a mammalian cell line for producing AAV as well as methods of using the mammalian cells for producing AAVs.

## Claims

### Claim 1

An isolated nucleic acid molecule operably comprising:
a. a first inducible promoter and a TetO2 sequence;
b. an E2A gene under control of the first inducible promoter;
c. a viral associated (VA) non-coding RNA under control of a second inducible promoter;
d. a third inducible promoter and a TetO2 sequence;
e. an E4 gene under control of the third inducible promoter;
f. an expression cassette encoding a protein ortholog of the VA RNA and a peptide protein tag under the control of a depressible promoter comprising a human Cytomegalovirus (CMV) promoter and a TetO2 sequence;
g. a termination sequence; and
h. an antibiotic resistance gene.

### Claim 2 (depends on 1)

The isolated nucleic acid molecule of claim 1, wherein the first inducible promoter is native to the E2A gene.

### Claim 3 (depends on 1)

The nucleic acid molecule of claim 1, wherein the second inducible promoter is a promoter native to the VA non-coding RNA.

### Claim 4 (depends on 1)

The nucleic acid molecule of claim 1, wherein the second inducible promoter is an H1 promoter.

### Claim 5 (depends on 1)

The isolated nucleic acid molecule of claim 1, wherein the third inducible promoter is native to the E4 gene.

### Claim 6 (depends on 1)

The isolated nucleic acid molecule of claim 1, wherein the protein ortholog is an influenza non-structural 1 protein (NS1), Ebola viral protein 35 (VP35), orthoreovirus sigma 3 protein (σ3), group C rotavirus non-structural protein 3 (NSP3), vaccinia virus E3L protein (E3L), herpes simplex virus type 1 US11 protein (US11), Epstein-Barr virus SM protein (SM), baculovirus PK2 protein (PK2), hepatitis C virus non-structural protein 5A protein (NS5A), human herpes virus-8 protein vIRF-2 (vIRF-2), human immunodeficiency virus protein Tat, vaccinia virus K3L protein (K3L), Herpes Simplex virus protein γ134.5/ICP34.5, and human papilloma virus-18 E6 protein (E6).

### Claim 7 (depends on 1)

The nucleic acid molecule of claim 1, wherein the nucleic acid sequence comprises a core insulator sequence inserted between the E4 gene and the expression cassette.

### Claim 8 (depends on 1)

The nucleic acid molecule of claim 1, wherein the nucleic acid sequence is flanked on both the 5′ and 3′ ends by transposon-specific inverted terminal repeats (ITR), and wherein the nucleic acid sequence comprises a core insulator sequence inserted downstream of the ITR on the 3′ end and upstream of the ITR on the 5′ end.

### Claim 9

An isolated nucleic acid molecule operably comprising a plasmid encoding:
a. a first inducible promoter and a TetO2 sequence;
b. a Rep78 gene, including a silenced p19 promoter located within the Rep78 gene coding region, wherein the silenced p19 promoter comprises mutations in SP1, TATA-1, and/or TATA-2 sites;
c. a second inducible promoter and a TetO2 sequence;
d. a Rep52 gene under control of the second inducible promoter;
e. termination sequence and;
f. an antibiotic resistance gene.

### Claim 10 (depends on 9)

The isolated nucleic acid molecule of claim 9, wherein the first inducible promoter is human cytomegalovirus (CMV) immediate early enhancer-promoter, CMV immediate early enhancer fused chicken β-actin promoter (CAG), human ubiquitin C (UbC) promoter or Rous sarcoma virus (RSV) promoter.

### Claim 11 (depends on 9)

The isolated nucleic acid molecule of claim 9, wherein the second inducible promoter is human CMV immediate early enhancer-promoter, CMV immediate early enhancer fused chicken β-actin promoter (CAG), human ubiquitin C (UbC) promoter or Rous sarcoma virus (RSV) promoter.

### Claim 12 (depends on 9)

The nucleic acid molecule of claim 9, wherein the nucleic acid sequence comprises a core insulator sequence inserted between the termination sequence and the antibiotic resistance gene.

### Claim 13 (depends on 9)

The nucleic acid molecule of claim 9, wherein the nucleic acid sequence is flanked on both the 5′ and 3′ ends by transposon-specific inverted terminal repeats (ITR), and wherein the nucleic acid sequence comprises a core insulator sequence inserted downstream of the ITR on the 3′ end and upstream of the ITR on the 5′ end.

### Claim 14

An isolated nucleic acid molecule operably comprising:
a. a first inducible promoter and a TetO2 sequence;
b. a VP1 gene;
c. a second inducible promoter and a TetO2 sequence;
d. a VP2 gene and a VP3 gene; and
e. an antibiotic resistance gene.

### Claim 15 (depends on 14)

The isolated nucleic acid molecule of claim 14, wherein the first inducible promoter is human cytomegalovirus (CMV) immediate early enhancer-promoter, CMV immediate early enhancer fused chicken β-actin promoter (CAG), human ubiquitin C (UbC) promoter or Rous sarcoma virus (RSV) promoter.

### Claim 16 (depends on 14)

The isolated nucleic acid molecule of claim 14, wherein the second inducible promoter is human CMV immediate early enhancer-promoter, CMV immediate early enhancer fused chicken β-actin promoter (CAG), human ubiquitin C (UbC) promoter or Rous sarcoma virus (RSV) promoter.

### Claim 17 (depends on 14)

The isolated nucleic acid molecule of claim 14, further comprising a gene of interest (GOI) downstream of the VP3 gene and upstream of the antibiotic resistance gene.

### Claim 18 (depends on 17)

The isolated nucleic acid molecule of claim 17, wherein the nucleic acid molecule comprises a core insulator sequence inserted between the VP3 gene and the GOI and between the GOI and the antibiotic resistance gene.

### Claim 19 (depends on 14)

The nucleic acid molecule of claim 14, wherein the nucleic acid sequence comprises a core insulator sequence inserted between the VP1 gene and the second inducible promoter and TetO2 sequence.

### Claim 20 (depends on 14)

The nucleic acid molecule of claim 14, wherein the nucleic acid sequence comprises a core insulator sequence inserted between the VP2 and VP3 gene of d) and the antibiotic resistance gene of e).

### Claim 21 (depends on 14)

The nucleic acid molecule of claim 14, wherein the nucleic acid sequence is flanked on both the 5′ and 3′ ends by transposon-specific inverted terminal repeats (ITR), and wherein the nucleic acid sequence comprises a core insulator sequence inserted downstream of the ITR on the 3′ end and upstream of the ITR on the 5′ end.