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NZ735329B2 - Optimized rpe65 promoter and coding sequences - Google Patents

Optimized rpe65 promoter and coding sequences Download PDF

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Publication number
NZ735329B2
NZ735329B2 NZ735329A NZ73532916A NZ735329B2 NZ 735329 B2 NZ735329 B2 NZ 735329B2 NZ 735329 A NZ735329 A NZ 735329A NZ 73532916 A NZ73532916 A NZ 73532916A NZ 735329 B2 NZ735329 B2 NZ 735329B2
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NZ
New Zealand
Prior art keywords
sequence
seq
rpe
nucleotides
vector
Prior art date
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NZ735329A
Other versions
NZ735329A (en
Inventor
Robin Ali
Alexander Smith
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Ucl Business Ltd
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Publication date
Priority claimed from GBGB1502137.1A external-priority patent/GB201502137D0/en
Application filed by Ucl Business Ltd filed Critical Ucl Business Ltd
Publication of NZ735329A publication Critical patent/NZ735329A/en
Publication of NZ735329B2 publication Critical patent/NZ735329B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
    • A61K48/0058—Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
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    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C12N15/861—Adenoviral vectors
    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
    • C12N2015/8518—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic expressing industrially exogenous proteins, e.g. for pharmaceutical use, human insulin, blood factors, immunoglobulins, pseudoparticles
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    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
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    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/008—Vector systems having a special element relevant for transcription cell type or tissue specific enhancer/promoter combination
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/01—Carboxylic ester hydrolases (3.1.1)
    • C12Y301/01064—Retinoid isomerohydrolase (3.1.1.64)

Abstract

The present invention relates to expression constructs comprising a retinal pigment epithelium (RPE) specific promoter and an operably linked polynucleotide sequence. The invention also relates to RPE specific promoters consisting of no more than 800 nucleotides. The present invention also relates to the prevention and/or treatment of retinal dystrophy in a patient, including Leber congenital amaurosis (LCA).

Claims (27)

1. An expression construct comprising a retinal pigment epithelium (RPE)-specific promoter and an operably linked polynucleotide sequence, wherein the RPE-specific 5 promoter comprises: (a) a sequence of contiguous nucleotides from SEQ ID NO: 1 that confers RPE-specific expression on an operably linked polynucleotide sequence, or (b) a sequence having at least 90% sequence identity to said sequence of (a) and that retains RPE-specific promoter activity, 10 and wherein the operably linked polynucleotide sequence comprises the sequence of SEQ ID NO: 4.
2. The expression construct of claim 1, wherein the RPE-specific promoter comprises: (a) no more than 1300 contiguous nucleotides from SEQ ID NO: 1, or 15 (b) a sequence having at least 90% sequence identity to said sequence of no more than 1300 contiguous nucleotides from SEQ ID NO:1 and retaining RPE-specific promoter activity.
3. The expression construct of claim 2, wherein the RPE-specific promoter comprises: (a) no more than 800 contiguous nucleotides from SEQ ID NO: 1, or 20 (b) a sequence having at least 90% sequence identity to said sequence of no more than 800 contiguous nucleotides from SEQ ID NO: 1 and retaining RPE-specific promoter activity.
4. The expression construct of claim 3, wherein the RPE-specific promoter comprises: (a) the sequence of SEQ ID NO: 2 or the sequence of nucleotides 12-761 of SEQ ID NO: 2, 25 or (b) a sequence having at least 90% sequence identity to said sequence of SEQ ID NO: 2, or to nucleotides 12-761 of SEQ ID NO: 2, and retaining RPE-specific promoter activity.
5. The expression construct according to claim 3, wherein the RPE-specific promoter 30 comprises SEQ ID NO: 2.
6. The expression construct according to claim 3, wherein the RPE-specific promoter consists of nucleotides 12-761 of SEQ ID NO: 2.
7. The expression construct according to any one of the preceding claims, wherein said 5 sequence of (a) or (b) is at least 700 nucleotides in length.
8. A retinal pigment epithelium (RPE)-specific promoter which consists of: (a) a sequence of no more than 800 contiguous nucleotides from SEQ ID NO:1 comprising nucleotides 12-761 of SEQ ID NO: 2, or 10 (b) a sequence having at least 90% sequence identity to nucleotides 12-761 of SEQ ID NO:2, and retaining RPE-specific promoter activity, wherein said sequence of (b) consists of no more than 800 nucleotides.
9. A retinal pigment epithelium (RPE)-specific promoter consisting of no more than 800 15 nucleotides, which comprises: (a) the sequence of SEQ ID NO: 2, or (b) a sequence having at least 90% sequence identity to said sequence of SEQ ID NO: 2, and retaining RPE-specific promoter activity. 20 10. The promoter of claim 9, wherein said promoter comprises the sequence of SEQ ID
10. NO: 2.
11. The promoter of claim 8, wherein said sequence consists of the sequence of nucleotides 12-761 of SEQ ID NO: 2, or a sequence having at least 90% sequence identity to 25 nucleotides 12-761 of SEQ ID NO: 2.
12. The promoter of claim 8 or 9, wherein said promoter of (b) consists of SEQ ID NO: 2, or a sequence having at least 90% sequence identity to SEQ ID NO: 2. 30
13. The promoter of any one of claims 8, 9, 11 or 12, wherein said sequence of (b) is at least 700 nucleotides in length.
14. A vector comprising an expression construct according to any one of claims 1 to 7 or a promoter according to any one of claims 8 to 13.
15. The vector according to claim 14, wherein the vector is a viral vector.
16. The vector according to claim 15, wherein the vector is an adeno-associated virus (AAV) vector or comprises an AAV genome or a derivative thereof.
17. The vector according to claim 16, wherein said derivative is a chimeric, shuffled or 10 capsid modified derivative.
18. The vector according to claims 16 or 17, wherein said AAV genome is from a naturally derived serotype or isolate or clade of AAV. 15 19. The vector according to claim 18, wherein said AAV genome is from AAV serotype 2
19. (AAV2), AAV serotype 4 (AAV4), AAV serotype 5 (AAV5) or AAV serotype 8 (AAV8) and/or wherein the capsid is derived from AAV5 or AAV8.
20. The vector according to claim 19, wherein the genome is derived from AAV2 and the 20 capsid is derived from AAV5 or AAV8.
21. An isolated host cell that contains a vector of claim 14 or produces a viral vector of any one of claims 15 to 20. 25
22. The cell according to claim 21 that is a HEK293 or HEK293T cell.
23. A pharmaceutical composition comprising a vector of any one of claims 14 to 20 and a pharmaceutically acceptable carrier. 30
24. Use of a vector according to any one of claims 14 to 20 in the manufacture of a medicament for the treatment or prevention of retinal dystrophy.
25. The use of claim 24, wherein the retinal dystrophy is Leber congenital amaurosis
26. (LCA), age-related macular degeneration (AMD), oculo-cutaneous type 1 or Nettleship-Falls type ocular albinism or MERTK deficiency. 5 26. The use according to claim 24 or 25, wherein the treatment or prevention is by administration of the vector according to any one of claims 14 to 20 to a patient by direct retinal, subretinal or intravitreal injection.
27. The use according to claim 26, wherein said vector is to be administered directly into 10 the retinal, subretinal space or intravitreal space. 1 2 3 4 1: AAV
NZ735329A 2016-02-08 Optimized rpe65 promoter and coding sequences NZ735329B2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1502137.1A GB201502137D0 (en) 2015-02-09 2015-02-09 Treatment
PCT/GB2016/050289 WO2016128722A1 (en) 2015-02-09 2016-02-08 Optimized rpe65 promoter and coding sequences

Publications (2)

Publication Number Publication Date
NZ735329A NZ735329A (en) 2024-08-30
NZ735329B2 true NZ735329B2 (en) 2024-12-03

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