scholarly journals Adeno-associated virus type 2-mediated transduction of murine hematopoietic cells with long-term repopulating ability and sustained expression of a human globin gene in vivo.

1997 ◽  
Vol 71 (4) ◽  
pp. 3098-3104 ◽  
Author(s):  
S Ponnazhagan ◽  
M C Yoder ◽  
A Srivastava
Keyword(s):  
Virus Type ◽  
Globin Gene ◽  
Hematopoietic Cells ◽  
Adeno Associated Virus

Induction of an Antitumoral Immune Response by Wild-Type Adeno-Associated Virus Type 2 in an In Vivo Model of Pancreatic Carcinoma

Pancreas ◽  
2007 ◽  
Vol 35 (1) ◽  
pp. 63-72 ◽  
Author(s):  
Sven Eisold ◽  
Jan Schmidt ◽  
Eduard Ryschich ◽  
Michael Gock ◽  
Ernst Klar ◽  
...  
Keyword(s):  
Immune Response ◽  
Pancreatic Carcinoma ◽  
Virus Type ◽  
In Vivo Model ◽  
Wild Type ◽  
Adeno Associated Virus

scholarly journals Conjugate-Based Targeting of Recombinant Adeno-Associated Virus Type 2 Vectors by Using Avidin-Linked Ligands

2002 ◽  
Vol 76 (24) ◽  
pp. 12900-12907 ◽  
Author(s):  
Selvarangan Ponnazhagan ◽  
Gandham Mahendra ◽  
Sanjay Kumar ◽  
John A. Thompson ◽  
Mark Castillas,
Keyword(s):  
Gene Therapy ◽  
Growth Factor ◽  
Virus Type ◽  
Transgene Expression ◽  
Future Application ◽  
Adeno Associated Virus ◽  
Tissue Specific ◽  
Prokaryotic Expression System

ABSTRACT The development of targeted vectors, capable of tissue-specific transduction, remains one of the important aspects of vector modification for gene therapy applications. Recombinant adeno-associated virus type 2 (rAAV-2)-based vectors are nonpathogenic, have relatively low immunogenicity, and are capable of long-term transgene expression. AAV-2 vectors bind primarily to heparan sulfate proteoglycan (HSPG), a receptor that is present in many tissues and cell types. Because of the widespread expression of HSPG on many tissues, targeted transduction in vivo appears to be limited with AAV-2 vectors. Thus, development of strategies to achieve transductional targeting will have a profound benefit in the future application of these vectors. We report here a novel conjugate-based targeting method to enhance tissue-specific transduction of AAV-2-based vectors. The present report utilized a high-affinity biotin-avidin interaction as a molecular bridge to cross-link purified targeting ligands, produced genetically as fusion proteins to core-streptavidin, in a prokaryotic expression system. Conjugation of the bispecific targeting protein to the vector was achieved by biotinylating purified rAAV-2 without abolishing the capsid structure, internalization, and subsequent transgene expression. The tropism-modified vectors, targeted via epidermal growth factor receptor (EGFR) or fibroblast growth factor 1α receptor (FGFR1α), resulted in a significant increase in transduction efficiency of EGFR-positive SKOV3.ip1 cells and FGFR1α-positive M07e cells, respectively. Further optimization of this method of targeting should enhance the potential of AAV-2 vectors in ex vivo and in vivo gene therapy and may form the basis for developing targeting methods for other AAV serotype capsids.

scholarly journals Adeno-Associated Virus Type 5 (AAV5) but Not AAV2 Binds to the Apical Surfaces of Airway Epithelia and Facilitates Gene Transfer

2000 ◽  
Vol 74 (8) ◽  
pp. 3852-3858 ◽  
Author(s):  
Joseph Zabner ◽  
Michael Seiler ◽  
Robert Walters ◽  
Robert M. Kotin ◽  
Wendy Fulgeras ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Virus Type ◽  
Target Cells ◽  
Apical Surface ◽  
Airway Epithelia ◽  
Adeno Associated Virus ◽  
Human Airway ◽  
Human Airway Epithelia

ABSTRACT In the genetic disease cystic fibrosis, recombinant adeno-associated virus type 2 (AAV2) is being investigated as a vector to transfer CFTR cDNA to airway epithelia. However, earlier work has shown that the apical surface of human airway epithelia is resistant to infection by AAV2, presumably as a result of a lack of heparan sulfate proteoglycans on the apical surface. This inefficiency can be overcome by increasing the amount of vector or by increasing the incubation time. However, these interventions are not very practical for translation into a therapeutic airway-directed vector. Therefore, we examined the efficiency of other AAV serotypes at infecting human airway epithelia. When applied at low multiplicity of infection to the apical surface of differentiated airway epithelia we found that a recombinant AAV5 bound and mediated gene transfer 50-fold more efficiently than AAV2. Furthermore, in contrast to AAV2, AAV5-mediated gene transfer was not inhibited by soluble heparin. Recombinant AAV5 was also more efficient than AAV2 in transferring β-galactosidase cDNA to murine airway and alveolar epithelia in vivo. These data suggest that AAV5-derived vectors bind and mediate gene transfer to human and murine airway epithelia, and the tropism of AAV5 may be useful to target cells that are not permissive for AAV2.

Enhanced sensitivity of pancreatic tumours cells to 5-FU-chemotherapy mediated by adeno-associated virus type 2 (AAV-2) infection in vitro and in vivo

2000 ◽  
Vol 118 (4) ◽  
pp. A531
Author(s):  
Sven Christian Eisold ◽  
Ruediger Ridder ◽  
Eduard Ryschisch ◽  
Jan Schmidt ◽  
Geeske C. Meyer ◽  
...  
Keyword(s):  
Virus Type ◽  
Adeno Associated Virus ◽  
Pancreatic Tumours ◽  
Enhanced Sensitivity

scholarly journals Relative Influence of the Adeno-Associated Virus (AAV) Type 2 p5 Element for Recombinant AAV Vector Site-Specific Integration

2007 ◽  
Vol 82 (5) ◽  
pp. 2590-2593 ◽  
Author(s):  
Mickaël Guilbaud ◽  
Gilliane Chadeuf ◽  
Fabio Avolio ◽  
Achille François ◽  
Philippe Moullier ◽  
...  
Keyword(s):  
Human Chromosome ◽  
Virus Type ◽  
Promoter Region ◽  
Raav Vector ◽  
Adeno Associated Virus ◽  
Site Specific ◽  
Site Specific Integration ◽  
Human Chromosome 19

ABSTRACT The p5 promoter region of the adeno-associated virus type 2 (AAV-2) rep gene has been described as essential for Rep-mediated site-specific integration (RMSSI) of plasmid sequences in human chromosome 19. We report here that insertion of a full-length or minimal p5 element between the viral inverted terminal repeats does not significantly increase RMSSI of a recombinant AAV (rAAV) vector after infection of growth-arrested or proliferating human cells. This result suggests that the p5 element may not improve RMSSI of rAAV vectors in vivo.

scholarly journals Adeno-Associated Virus Type 2 Capsids with Externalized VP1/VP2 Trafficking Domains Are Generated prior to Passage through the Cytoplasm and Are Maintained until Uncoating Occurs in the Nucleus

2006 ◽  
Vol 80 (22) ◽  
pp. 11040-11054 ◽  
Author(s):  
Florian Sonntag ◽  
Svenja Bleker ◽  
Barbara Leuchs ◽  
Roger Fischer ◽  
Jürgen A. Kleinschmidt
Keyword(s):  
Virus Type ◽  
Catalytic Domain ◽  
Nuclear Translocation ◽  
Basic Amino Acid ◽  
Nuclear Entry ◽  
Direct Role ◽  
Adeno Associated Virus

ABSTRACT Common features of parvovirus capsids are open pores at the fivefold symmetry axes that traverse the virion shell. Upon limited heat treatment in vitro, the pores can function as portals to externalize VP1/VP2 protein N-terminal sequences which harbor infection-relevant functional domains, such as a phospholipase A2 catalytic domain. Here we show that adeno-associated virus type 2 (AAV2) also exposes its VP1/VP2 N termini in vivo during infection, presumably in the endosomal compartment. This conformational change is influenced by treatment with lysosomotropic reagents. While incubation of cells with bafilomycin A1 reduced exposure of VP1/VP2 N termini, incubation with chloroquine stimulated externalization transiently. N-terminally located basic amino acid clusters with nuclear localization activity also become exposed in this process and are accessible on the virus capsid when it enters the cytoplasm. This is an obligatory step in AAV2 infection. However, a direct role of these sequences in nuclear translocation of viral capsids could not be determined by microinjection of wild-type or mutant viruses. This suggests that further modifications of the capsid have to take place in a precytoplasmic entry step that prepares the virus for nuclear entry. Microinjection of several capsid-specific antibodies into the cell nucleus blocked AAV2 infection completely, supporting the conclusion that AAV2 capsids bring the infectious genome into the nucleus.

scholarly journals The Cellular TATA Binding Protein Is Required for Rep-Dependent Replication of a Minimal Adeno-Associated Virus Type 2 p5 Element

2005 ◽  
Vol 79 (17) ◽  
pp. 11082-11094 ◽  
Author(s):  
Achille François ◽  
Mickaël Guilbaud ◽  
Rafi Awedikian ◽  
Gilliane Chadeuf ◽  
Philippe Moullier ◽  
...  
Keyword(s):  
Virus Type ◽  
Transcription Initiation ◽  
Promoter Region ◽  
Binding Protein ◽  
Tata Binding Protein ◽  
Tata Box ◽  
Adeno Associated Virus

ABSTRACT The p5 promoter region of adeno-associated virus type 2 (AAV-2) is a multifunctional element involved in rep gene expression, Rep-dependent replication, and site-specific integration. We initially characterized a 350-bp p5 region by its ability to behave like a cis-acting replication element in the presence of Rep proteins and adenoviral factors. The objective of this study was to define the minimal elements within the p5 region required for Rep-dependent replication. Assays performed in transfected cells (in vivo) indicated that the minimal p5 element was composed by a 55-bp sequence (nucleotides 250 to 304 of wild-type AAV-2) containing the TATA box, the Rep binding site, the terminal resolution site present at the transcription initiation site (trs+1), and a downstream 17-bp region that could potentially form a hairpin structure localizing the trs+1 at the top of the loop. Interestingly, the TATA box was absolutely required for in vivo but dispensable for in vitro, i.e., cell-free, replication. We also demonstrated that Rep binding and nicking at the trs+1 was enhanced in the presence of the cellular TATA binding protein, and that overexpression of this cellular factor increased in vivo replication of the minimal p5 element. Together, these studies identified the minimal replication origin present within the AAV-2 p5 promoter region and demonstrated for the first time the involvement of the TATA box, in cis, and of the TATA binding protein, in trans, for Rep-dependent replication of this viral element.

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