WO2009125902A1 - Vecteur à base d'un aav (adeno-associated virus) sérotype 5 pour la délivrance ciblée d'un gène - Google Patents

Vecteur à base d'un aav (adeno-associated virus) sérotype 5 pour la délivrance ciblée d'un gène Download PDF

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WO2009125902A1
WO2009125902A1 PCT/KR2008/004476 KR2008004476W WO2009125902A1 WO 2009125902 A1 WO2009125902 A1 WO 2009125902A1 KR 2008004476 W KR2008004476 W KR 2008004476W WO 2009125902 A1 WO2009125902 A1 WO 2009125902A1
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seq
amino acid
virus
vector
cells
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Heui Ran Lee
Han Seok Choi
Han Sam Lee
Jee Yeon Kim
Oh Kyu Shin
Won Il Lee
Sun Joo Jung
Kee Rang Park
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University of Ulsan Foundation for Industry Cooperation
Industry Academic Cooperation Foundation of Dankook University
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Industry Academic Cooperation Foundation of Dankook University
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    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
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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
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    • 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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    • 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
    • C12N2750/14145—Special targeting system for viral vectors
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    • C12N2810/00—Vectors comprising a targeting moiety
    • C12N2810/40—Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source
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    • C12N2810/00—Vectors comprising a targeting moiety
    • C12N2810/40—Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source
    • C12N2810/405—Vectors comprising RGD peptide

Definitions

  • the present invention relates to a targetable, adeno-associated virus serotype
  • 5-derived recombinant virus vector prepared by inserting a targeting motif peptide, specifically recognizing a target molecule, into the internal amino acid residue of a capsid protein, and to a method of delivering a gene in a target cell-specific manner using said vector.
  • Recombinant adeno-associated virus possesses many properties that make an attractive vector to deliver a gene of interest.
  • rAAV capable of infecting both dividing and non-dividing cells, offers persistent transgene expression with induction of minimal immune response [Wu, Z. et al., 2006, MoI Ther 14: 316-327; Warrington, K. H. et al., 2006, Hum Genet 1 19: 571 -603].
  • AAV serotypes which can be used to delivery target genes, the most widely studied vector is an adeno-associated virus serotype 2, which is now being used for clinical gene transfer for cystic fibrosis [Moss, R.
  • rAAV 1 serotype there is an example in which an RGD-4C motif was inserted at amino acid 590 of rAAV l capsid protein corresponding to amino acid 587 of rAAV2 capsid in order to target vascular endothelial cells and to alter tropism [MD Stachler, MD. et al., 2006, Gene Therapy 13:926-931 ].
  • RGD-4C motif was inserted at amino acid 590 of rAAV l capsid protein corresponding to amino acid 587 of rAAV2 capsid in order to target vascular endothelial cells and to alter tropism
  • the data of the present invention suggested that the introduction of homing peptide at amino acid 578 sharply drops TE by rAAV5, probably knocking down innate rAAV5 tropism. Thus, a region near amino acid 578 residing on threefold-proximal peak is likely responsible for rAAV5 binding to sialic acid, not the region on the inward surface within twofold axis. However, more study has to be carried out to elucidate the detailed features for the receptor binding phenotype of rAAV5.
  • Alpha V integrins are often over-expressed in a wide variety of tumor cells and vasculature [Bello, L., et al., 2001 , Neurosurgery 49: 380-389; discussion 390; Brooks, P. C, et al., 1994, Science 264: 569-571].
  • Alpha V integrin-binding RGD motif (RGD-4C) was initially introduced to genetically targeting adenovirus and this mutant significantly enhanced human cancer cells both in vitro and in vivo [Kanerva, A., et al., 2004, International journal of cancer 110: 475-480; Niu, G., et al., 2007, MoI Imaging Biol].
  • rAAV2 exposing RGD motif was successfully generated to retarget rAAV2 IGirod, A., et al. 1999, Nat Med 5: 1438; Shi, W., etal., 2006, Gynecologic oncology 103: 1054-1062].
  • Salyl Lewis X (sLeX) is found on the surfaces of various cancer cells and the sLeX binding motif was previously identified by peptide - AHWIPRYSSPAT- binding analysis [Kwon, M., et al., 2002, J Am Chem Soc 124: 13996-13997].
  • Tenascin C (TnC) is frequently over-expressed in different human tumors [Wiksten, J. P., et al., 2003, Oncology 64: 245-250]. TnC-homing peptide was identified by the phage display as the same strategy for sLeX.
  • rAAV has been identified as a proper vector to obtain long-term anti-tumoral effect [Davidoff, A. M., et al., 2002, Cancer research 62: 3077-3083].
  • systemic administration of rAAV in vivo results in the localization of the majority of the virus in the liver and spleen, thus limiting its application [Nathwani, A. C, et al., 2007, Blood 109: 1414-14211.
  • Baker and co- workers accomplished targeted delivery to vascular tissue by rAAV2 that possess the desired tropism [Work, L. M., et al., 2006, MoI Ther 13: 683-693; White, K., et al., 2007, Gene therqy White, S. J., et al. 2004, Circulation 109: 513-519].
  • Transduction efficiency (TE) mediated by rAAV would be negatively influenced by pre-existing neutralizing antibodies.
  • TE Transduction efficiency
  • the potency of rAAV2 as a gene delivery vehicle would be severely hindered for in vivo application, particularly in case of systemic administration of rAAV2.
  • a capsid protein constituting the envelope of adeno-associated virus serotype 5 were modified such that the virus could target specific cells.
  • a vector producing a virus which has modified surface characteristics and can recognize and bind to a target molecule, was constructed by inserting a targeting motif peptide into the capsid protein of a virus vector.
  • the present invention relates to a targeted recombinant virus vector derived from adeno-associated virus serotype 5, in which the virus vector comprises a nucleic acid sequence encoding a capsid protein containing a targeting motif peptide inserted in the internal amino acid residue of the capsid protein.
  • the virus vector of the present invention is an adeno-associated virus serotype
  • the virus vector can deliver a therapeutic gene or a genetic substance specifically into cells, tissues and/or organs.
  • the vector of the present invention expresses a chimeric capsid protein, in which a targeting motif peptide is linked to a capsid protein, a viral structural protein, in the form of a single- stranded polypeptide through a peptide bond.
  • a virus produced from the vector of the present invention has an envelope having tropism specific for targets depending on the kind and characteristics of a peptide, inserted therein, and it also has the capability to transduce cells depending on the kind and characteristics of peptide.
  • the tropism and characteristics of the vector of the present invention can be easily manipulated according to the intended use thereof by controlling the kind of a peptide which is inserted into the vector.
  • a pep tide-encoding nucleic acid sequence is inserted in a capsid protein-encoding nucleic acid sequence, such that the capsid protein, recombined at the genetic level, and a motif peptide, are expressed as one polypeptide.
  • targeting motif peptide refers to a peptide capable of specifically recognizing and binding to a target molecule, which is present specifically in the membrane of specific cells or present in the membrane in excess.
  • Targeting motif peptides that can be inserted into the vector of the present invention include all peptides which can impart a targeting function to virus without losing the virus- producing ability.
  • the targeting motif peptide of the present invention is a peptide specifically recognizing and/or binding to molecules, which are expressed in tumor cells, for example, sialic Lewis X, integrin or Tenascin C. More specifically, the targeting motif peptide specifically recognizing sialic Lewis X has an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, the motif peptide specifically recognizing integrin has an amino acid sequence of SQ ID NO: 3 or SEQ ID NO: 4, and the motif peptide specifically recognizing Tenascin C has an amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8.
  • the size of the targeting motif peptide is limited to a size which can produce virus without losing the viral replication ability and assembly ability, when the peptide is inserted into the vector and produced as virus.
  • the targeting motif peptide preferably comprises 5-25 amino acids, and more preferably 7-20 amino acids. Peptide sequences having various sizes ranging from 7 to 20 amino acids could be readily inserted into a capsid protein without losing the viral replication and assembly ability, while they have particular buoyant density. There was no mutual relation between insert size and the efficiency of producing virus particles within this range. This size in the present invention is much longer than previously determined in case of rAAV2 bearing 17 amino acids [Blacklow, N. R., et al., 1968, Journal of the National Cancer Institute 40: 319-327].
  • the motif peptide of the present invention is inserted at amino acid residues of a capsid protein which can be genetically modified without affecting the viral replication ability and assembly ability.
  • the present inventors have found that both loops of amino acids 444 and 578 of adeno-associated virus serotype 5 are regions which can be genetically modified without affecting the viral replication ability and assembly ability.
  • a recombinant adeno-associated virus serotype 5 can be genetically modified within the capsid region, particularly at VPl amino acid position 444 or 578, into which an external peptide can be inserted; ii) mutants having inserted therein the targeting motif peptide having the above-specified size have a virus production ability equal or similar to that of wild- type virus; and iii) viruses genetically modified by the insertion of the peptide can transduce human cancer cells depending on the characteristics of retargeted receptor.
  • rAAV5 recombinant adeno-associated virus serotype 5
  • vector of the present invention can be amended, modified or applied according to any conventional method known in the vector preparation field.
  • the vector of the present invention can be used for the prevention and treatment of various diseases in the medical and veterinary fields by inserting a nucleic acid, encoding at least one gene to be delivered and expressed in cells, tissues and organs, into the vector.
  • genes which can be inserted into the vector of the present invention, include, but are not limited to, various physiologically active peptides, such as hormones, cytokines, enzymes, antibodies, growth factors, transcription regilatory factors, blood factors, vaccines, structural proteins, drug-sensitive genes, drug-resistant genes, ligand proteins and receptors, cell surface antigens, receptor antagonists, and derivatives thereof.
  • they may include anti-sense nucleic acid regilating gene expression in target cells, or nucleic acid expressing siRNA.
  • the present invention relates to a method of delivering a gene in a target cell-specific manner using a targeted recombinant virus vector, which is derived from adeno-associated virus serotype 5 and contains a nucleic acid sequence encoding a capsid protein comprising a targeting motif peptide inserted into the amino acid sequence thereof.
  • the inventive targeted recombinant virus vector encoding at least one gene to be delivered or expressed in cells, tissues and/or organs can proliferate into viruses using adeno-associated virus replication methods known in the art, for example, using helper virus such as adenovirus, vaccinia virus or herpes virus, in consideration of purity, stability, morphology and infectivity.
  • helper virus such as adenovirus, vaccinia virus or herpes virus
  • the adeno-associated virus serotype 5 virus can be used as a gene carrier in not only humans, but also animals having infectivity with virus, through an ex vivo method of transferring the virus into cells isolated from an individual and then transplanting the cells again into the individual, or an in vivo method of injecting the virus directly into an individual.
  • the vector and virus of the present invention can be used to treat various diseases, including blood diseases, such as hemophilia, sickle cell anemia, and anemia caused by chemotherapy, central nervous system diseases, such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, Huntington's disease, and mental disorder caused by genetic defect, metabolic diseases, such as cystic fibrosis, diabetes, growth hormone deficiency and osteoporosis, autoimmune diseases, such as multiple sclerosis, psoriasis and rheumatoid disease, and infectious diseases, such as HlV infection, influenza, herpes simplex, papillomavirus and mycobacteria.
  • blood diseases such as hemophilia, sickle cell anemia, and anemia caused by chemotherapy
  • central nervous system diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, Huntington's disease, and mental disorder caused by genetic defect
  • metabolic diseases such as cystic fibrosis, diabetes, growth hormone deficiency and osteoporosis
  • the virus of the present invention may be formulated into a suitable preparation form together with a pharmaceutically acceptable carrier and administered via various routes, for example, oral, intraperitoneal, intravenous, intramuscular, subcutaneous, intracutaneous, local, intranasal, intrapulmonary and intrarectal routes.
  • routes for example, oral, intraperitoneal, intravenous, intramuscular, subcutaneous, intracutaneous, local, intranasal, intrapulmonary and intrarectal routes.
  • the adeno-associated virus serotype 5 mutants produced from the vector of the present invention showed a virus-producing ability similar to that of wild-type virus without losing the virus production ability, and showed transducing activity specific for certain cells depending on the kind and characteristics of a peptide inserted therein.
  • FlG. 1 shows the structure of the mutant AAV5 capsid subunit and the topology of the virus surface.
  • FlG. l(a) shows the schematic structure of a pSp72-R2C5 plasmid containing AAV2 rep coding gene (REP) and VPl capsid coding gene (CAP)
  • FIG. l (b) shows the surface of rAAV5-sLeXl at aa 444 site (red) and rAAV5-sLeX2 at aa 578 (yellow) viewed as ribbons
  • FIG. l (c) shows the comparison of the backbones of AAV2 (blue) and rAAV5-RGDl (green).
  • FIG. l(a) shows the schematic structure of a pSp72-R2C5 plasmid containing AAV2 rep coding gene (REP) and VPl capsid coding gene (CAP)
  • FIG. l (b) shows the surface of rA
  • FIG. l(c) illustrates the red loop region is an RGD-4C peptide domain inserted at aa578 site. This area is magnified and viewed as the colorful spheres and stick forms.
  • FIG. l (d) illustrates the cartoon of rAAV5-RGDl, which is assembled by 60 capsid subunits with icosahedral symmetry. In FIG. l (d), all subsequent surface rendering was performed using PyMOL, and red fragments show RGD-based homing motives at aa 578 position.
  • FIG. 2 shows transmission electron microscopy pictures of rAAV5 and rAAV5-sLeX2 viruses. Specifically, FIG. 2(a) shows wild-type rAAV5 particles, and FIG. 2(b) shows mutant rAAV5-sLeX2 particles. In FIG. 2, all the virus particles were in 1 OmM Tris buffer (pH7.9) containing 2 inM MgCl 2 and 2% sorbitol.
  • FIG. 3 shows RGDS peptide-mediated inhibition of rAAV5-RGD 1 transduction in integrin-positive cancer cells.
  • HeLa cells, SK-Hep l cells, and U87-MG cells were infected with rAAV5-eGFP (MOI 10, 100, and 200, respectively), or rAAV5-RGDl (MOI 5,000, 10,000, and 5,000), respectively. These cells were incubated with RGDS or non-specific RGES peptide.
  • FIG. 3(a) shows the results of flow cytometry of GFP-positive cells after 48 hr
  • human cancer cells hepatocellular SK-Hep l , cervical HeLa, breast MDA-MB-231 and MDA-MD-435S, glioblastoma U-251, U87-MG, Ul 18-MG, colon HCT-116) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). The cells were incubated in Dulbecco's modified Eagle's medium, supplemented with 10% fetal bovine serum, L-glutamax (2 mM), penicillin (100 IU/ml) and streptomycin (50 ⁇ g/ml), in a 5% CO 2 incubator at 37 0 C.
  • Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, L-glutamax (2 mM), penicillin (100 IU/ml) and streptomycin (50 ⁇ g/ml
  • the cells were treated with rAAVs at various multiplicities of infection (MOI).
  • MOI multiplicities of infection
  • Mock- treated cells used as a control were cells not treated with rAAVs.
  • GFP expression was observed under an inverted fluorescent microscope (Leica DMIRB, Leica, Germany) and the pictures were taken.
  • pR2C5 and pHpa-trs-SK were obtained from Dr. High, and the adenoviral pXX6 helper plasmid was obtained from Stratagene (La jolla, CA, USA).
  • pcDNA3.1(+)-sc39TK was provided by Donald B Kbhn (University of Southern California, Keck School of Medicine, University of Los Angeles).
  • pHpa-trs-SK was composed of the EGFP expression cassette in the self-complementary AAV2 (scAAV2) genome.
  • the pSp72-R2C5 plasmid was constructed by subcloning of the 4.2-kb partial Kpn ⁇ -Xba ⁇ fragment of pR2C5 into the Kpn ⁇ -Xba ⁇ sites of the pSp72 vector. Then, for easy subcloning, the production of SmBl site at 1210 bp to 1215 bp without replacement of codon in rAAV5 VPl-encoding sequence of pSp72-R2C5 was performed using the QiikChange Multi Ste-Drected Mutagenesis Mt as described by the supplier (Stratagene, La Jolla, California), and this plasmid was named pSp72-R2C5-5Ar ⁇ Bl. The 1.0-kb Xbal-SnaBl fragment of a PCR product was obtained from pSp72-R2C5 using primers of SEQ ID NO: 9 and SEQ ID NO: 10.
  • the plasmid pGEM-T easy-Cap5 was obtained by TA cloning the 1.0 kb Xbal-SnaBl fragment of pR2C5 into the 3' T overhangs at the insertion site of pGEM -T Easy Vector (Promega, Madison, WI).
  • a PCR fragment of the QiikChange Multi Ste-Drected Mutagenesis was prepared using the pGEM T-easy-Cap5 plasmid as a template with each of primer pairs: one (Forward) containing more than 15 nucleotides belonging to the VPl gene upstream of the insertion site and also some nucleotides coding for the 5' terminal end of the motive peptide, and the other (Reverse) containing the over 15 nucleotides belonging to the cap gene downstream of the insertion site and also some nucleotides coding for the 3' terminal end of the motive peptide.
  • the following primer pairs were used: SEQ ID NO: 1 1 and SEQ ID NO: 12, SEQ ID NO: 13 and SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18, SEQ ID NO: 19 and SEQ TD NO: 20, SEQ ID NO: 21 and SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, and SEQ ID NO: 25 and SEQ ID NO: 26.
  • the 1.0-kb TnC3 PCR fragment was obtained from the template pGEM-Cap5-TnC 1.
  • the PCR products were ligated, amplified into XL-Blue E. coli (Strategene, La Jolla, California) and sequenced.
  • pSp72-scAAV2-eGFP was constructed by blunt-end subcloning of the 2.3-kb PvuW- Pvull fragment of pHpa-trs-SK into the Bgfll and HmdIII sites of the pSp72 vector, and thus it contained the full-length self-complementary AAV2 genome encoding an EGFP protein.
  • pSp72-scAAV2-sc39TK was constructed by subcloning of the BamHl-Sall PCR fragment of sc39TK gene from pcDNA3.1(+)-sc39TK into the BamHl-Sall sites in pSp72-scAAV2-eGFP from which the corresponding fragment had been removed.
  • Pure populations of rAAVs were prepared by two sequential steps of CsCl gradient ultracen- trifugation. After dialysis against 50 mM Tris buffer (p ⁇ 7.8) containing 1 mM MgCl 2 and 10% sorbitol, the purified rAAVs were aliquoted and stored at -80 0 C.
  • the total number of rAAV particles was calculated by TaqMan-based real-time PCR analysis (iQTM supermix, ⁇ o-Rad, Hercules, CA), using primers of SEQ ID NO: 27 and SEQ ID NO: 28 and a primer of SEQ ID NO: 29 (the 5' end of the primer was linked with FAM, and the 3' end of the primer was linked with TAMRA) targeting the cytomegalovirus (CMV) promoter [Moon, M. S., et al., 2005, lntervirology 48: 153- 160].
  • CMV cytomegalovirus
  • HeLa. SK-Hepl and U87-MG cells were cultured in 48-well plates for 24 hr before treatment with the peptide. The cells were incubated in fresh media supplemented with 200, 100, 50, 20, 10 ⁇ M active RGDS (Sgma, Steinheim, Germany), and inactive RGES peptide (Sgma, Sgma, Steinheim, Germany) with 2% FBS for 30 min at room temperature. Then, the cells were treated with rAAV5-GFP or mutants containing GFP expressing cassette (virus titers at approximate 50% transduction) for 4 hr.
  • active RGDS Sgma, Steinheim, Germany
  • RGES peptide Sgma, Sgma, Steinheim, Germany
  • adenovirus type 5 PFU 2 or 5 per cell
  • the cells were washed three times with PBS. 48 hours after the transduction, the cells each from the treated and untreated groups were pelleted and resuspended in 200 ⁇ i PBS or 200 ⁇ Jt PBS containing 4% paraformaldehyde. GFP-positive cells were measured by a flow cytometer.
  • RNA was isolated from HCT- 1 16, U251 , U87-MG, and U l 18-MG using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instruction.
  • RT-PCR was performed using a TNfn 5 forward primer of SEQ ID NO: 30 and a TNfn Al reverse primer of SEQ ID NO: 31, and for the detection of the TNC C-terminal form expression, RT- PCR was performed using a TNfbg forward primer of SEQ ID NO: 32 and a TNfbg reverse primer of SEQ ID NO: 33.
  • the present inventors first analyzed the mutants for their ability to amplify virus particles and the data showed that virus production was mostly as efficient as that of wild-type rAAV5 (Table 1). Virus production rate ranged from 7.3 x 10 10 to 2.8 x 10" virus genome copies per ml. After treatment with nuclease benzonaze, virus particle remained intact without losing its transducing activity (data not shown). This indicates that the efficient virus assembly occurred and the resultant virus particles are intact as much as the parental type.
  • the buoyant densities of each virus under CsQ 2 gradient were within a narrow range from 1.380 to 1.385 g/ml, slightly lighter than that of the wild-type (1.394 ⁇ 0.007 g/ml).
  • VG viral genome. Genome copies were determined by real-time PCR as described in Materials and Methods. Relative titer shows the ratio of the mutant virus titer to wild rAAV5 titer. Buoyant density of CsCU gradients menas the CsCk density of the major peak of rAAV5 mutants vectors purified and fractioned. psj ⁇ Comparison of virus particle-procfccing ability between
  • the adeno-associated virus serotype 5-derived targetable vector of the present invention has excellent safety and transducing activity and can be widely used to prevent and treat various diseases, including blood diseases, central nervous system diseases, metabolic diseases, autoimmune diseases, infectious diseases and tumors, in the medical and veterinary fields.

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Abstract

Cette invention concerne l'utilisation à titre de vecteur génique d'un AAV (adeno-associated virus) sérotype 5 d'une sécurité élevée, parmi les sérotypes AAV. Plus spécifiquement, cette invention décrit un vecteur viral recombiné, pouvant être ciblé, dérivé d'un AAV sérotype 5, préparé par insertion d'un peptide à motif de ciblage, reconnaissant spécifiquement une molécule cible, dans le résidu acide aminé interne d'une protéine de capside, et un procédé de délivrance d'un gène de manière spécifique de cellule cible au moyen dudit vecteur.
PCT/KR2008/004476 2008-04-07 2008-08-01 Vecteur à base d'un aav (adeno-associated virus) sérotype 5 pour la délivrance ciblée d'un gène Ceased WO2009125902A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020080032245A KR100912270B1 (ko) 2008-04-07 2008-04-07 표적화된 유전자 전달을 위한 아데노-부속 바이러스 혈청형5 벡터
KR10-2008-0032245 2008-04-07

Publications (1)

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WO2009125902A1 true WO2009125902A1 (fr) 2009-10-15

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PCT/KR2008/004476 Ceased WO2009125902A1 (fr) 2008-04-07 2008-08-01 Vecteur à base d'un aav (adeno-associated virus) sérotype 5 pour la délivrance ciblée d'un gène

Country Status (2)

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KR (1) KR100912270B1 (fr)
WO (1) WO2009125902A1 (fr)

Cited By (1)

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WO2023103662A1 (fr) * 2021-12-10 2023-06-15 和元生物技术（上海）股份有限公司 Mutant de virus adéno-associé approprié pour une infection spécifique de cellules u87-mg

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Publication number Priority date Publication date Assignee Title
KR101154374B1 (ko) * 2009-02-18 2012-07-09 재단법인 아산사회복지재단 표적화된 유전자 전달을 위한 아데노-부속 바이러스 혈청형5 벡터를 포함하는 조성물

Non-Patent Citations (5)

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Title
DAVIDSON, B. L. ET AL.: "Recombinant adeno-associated virus type 2, 4, and 5 vectors: transduction of variant cell types and regions in the mammalian central nervous system.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, USA., vol. 97, no. 7, 28 March 2000 (2000-03-28), pages 3428 - 3432 *
GRIFMAN, M. ET AL.: "Incorporation of Tumor-targeting peptides into recombinant adeno-associated virus capsids.", MOLECULAR THERAPY., vol. 3, no. 6, June 2001 (2001-06-01), pages 964 - 975 *
HOLIG, P. ET AL.: "Novel RGD lipopeptides for the targeting ofliposomes to integrin-expressing endothelial and melanoma cells.", PROTEIN ENGINEERING, DESIGN & SELECTION., vol. 17, no. 5, 21 July 2004 (2004-07-21), pages 433 - 441 *
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023103662A1 (fr) * 2021-12-10 2023-06-15 和元生物技术（上海）股份有限公司 Mutant de virus adéno-associé approprié pour une infection spécifique de cellules u87-mg

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