World Library  
Flag as Inappropriate
Email this Article

Adeno-associated virus

Article Id: WHEBN0001883421
Reproduction Date:

Title: Adeno-associated virus  
Author: World Heritage Encyclopedia
Language: English
Subject: Gene therapy of the human retina, Provirus, Leber's congenital amaurosis, Gene therapy for osteoarthritis, Gene therapy
Collection: Applied Genetics, Experimental Cancer Treatments, Parvovirinae, Parvoviruses
Publisher: World Heritage Encyclopedia

Adeno-associated virus

Adeno-associated virus
Adeno-associated virus serotype 2 structure from 1LP3. One fivefold axis shown center.
Virus classification
Group: Group II (ssDNA)
Order: Unassigned
Family: Parvoviridae
Subfamily: Parvovirinae
Genus: Dependoparvovirus
  • Adeno-associated dependoparvovirus A
  • Adeno-associated dependoparvovirus B

Adeno-associated virus (AAV) is a small virus which infects humans and some other primate species. AAV is not currently known to cause disease. The virus causes a very mild immune response, lending further support to its apparent lack of pathogenicity. Gene therapy vectors using AAV can infect both dividing and quiescent cells and persist in an extrachromosomal state without integrating into the genome of the host cell, although in the native virus some integration of virally carried genes into the host genome does occur.[1] These features make AAV a very attractive candidate for creating viral vectors for gene therapy, and for the creation of isogenic human disease models.[2] Recent human clinical trials using AAV for gene therapy in the retina have shown promise.[3]

AAV belongs to the genus Dependoparvovirus, which in turn belongs to the family Parvoviridae. The virus is a small (20 nm) replication-defective, nonenveloped virus.


  • Advantages and drawbacks 1
  • Clinical trials 2
  • Pathology 3
  • AAV structure 4
    • AAV genome, transcriptome and proteome 4.1
      • ITR sequences 4.1.1
      • rep genes and Rep proteins 4.1.2
      • cap genes and VP proteins 4.1.3
  • AAV serotypes, receptors and native tropism 5
    • Serotype 2 5.1
      • Serotype 2 and cancer 5.1.1
    • Other Serotypes 5.2
  • AAV immunology 6
    • Innate 6.1
    • Humoral 6.2
    • Cell-mediated 6.3
  • AAV infection cycle 7
  • See also 8
  • References 9
  • External links 10

Advantages and drawbacks

Wild-type AAV has attracted considerable interest from gene therapy researchers due to a number of features. Chief amongst these is the virus's apparent lack of pathogenicity. It can also infect non-dividing cells and has the ability to stably integrate into the host cell genome at a specific site (designated AAVS1) in the human chromosome 19.[4][5] The feature makes it somewhat more predictable than retroviruses, which present the threat of a random insertion and of mutagenesis, which is sometimes followed by development of a cancer. The AAV genome integrates most frequently into the site mentioned, while random incorporations into the genome take place with a negligible frequency. Development of AAVs as gene therapy vectors, however, has eliminated this integrative capacity by removal of the rep and cap from the DNA of the vector. The desired gene together with a promoter to drive transcription of the gene is inserted between the inverted terminal repeats (ITR) that aid in concatamer formation in the nucleus after the single-stranded vector DNA is converted by host cell DNA polymerase complexes into double-stranded DNA. AAV-based gene therapy vectors form episomal concatemers in the host cell nucleus. In non-dividing cells, these concatemers remain intact for the life of the host cell. In dividing cells, AAV DNA is lost through cell division, since the episomal DNA is not replicated along with the host cell DNA. Random integration of AAV DNA into the host genome is detectable but occurs at very low frequency. AAVs also present very low immunogenicity, seemingly restricted to generation of neutralizing antibodies, while they induce no clearly defined cytotoxic response.[6][7][8] This feature, along with the ability to infect quiescent cells present their dominance over adenoviruses as vectors for human gene therapy.

Use of the virus does present some disadvantages. The cloning capacity of the vector is relatively limited and most therapeutic genes require the complete replacement of the virus's 4.8 kilobase genome. Large genes are, therefore, not suitable for use in a standard AAV vector. Options are currently being explored to overcome the limited coding capacity. The AAV ITRs of two genomes can anneal to form head to tail concatamers, almost doubling the capacity of the vector. Insertion of splice sites allows for the removal of the ITRs from the transcript.

Because of AAV's specialized gene therapy advantages, researchers have created an altered version of AAV termed self-complementary adeno-associated virus (scAAV). Whereas AAV packages a single strand of DNA and must wait for its second strand to be synthesized, scAAV packages two shorter strands that are complementary to each other. By avoiding second-strand synthesis, scAAV can express more quickly, although as a caveat, scAAV can only encode half of the already limited capacity of AAV.[9] Recent reports suggest that scAAV vectors are more immunogenic than single stranded adenovirus vectors, inducing a stronger activation of cytotoxic T lymphocytes.[10]

The humoral immunity instigated by infection with the wild type is thought to be a very common event. The associated neutralising activity limits the usefulness of the most commonly used serotype AAV2 in certain applications. Accordingly the majority of clinical trials currently underway involve delivery of AAV2 into the brain, a relatively immunologically privileged organ. In the brain, AAV2 is strongly neuron-specific.

Clinical trials

To date, AAV vectors have been used in over 117 clinical trials worldwide. (Approximately 5.6%)[11] Recently, promising results have been obtained from Phase 1 and Phase 2 trials for a number of diseases, including Leber's Congenital Amaurosis,[12][13][14] Hemophilia,[15] congestive heart failure,[16] lipoprotein lipase deficiency (reference needed), and Parkinson's disease.[17]

Selected Clinical Trials using AAV-Based Vectors[18]
Indication Gene Route of administration Phase Subject number Status
Cystic fibrosis CFTR Lung, via aerosol I 12 Complete
CFTR Lung, via aerosol II 38 Complete
CFTR Lung, via aerosol II 100 Complete
Hemophilia B FIX Intramuscular I 9 Complete
FIX Hepatic artery I 6 Ended
Arthritis TNFR:Fc Intraarticular I 1 Ongoing
Hereditary emphysema AAT Intramuscular I 12 Ongoing
Leber's Congenital Amaurosis RPE65 Subretinal I-II Multiple Several ongoing and complete
Age-Related Macular Degeneration sFlt-1 Subretinal I-II 24 Ongoing
Muscular dystrophy Sarcoglycan Intramuscular I 10 Ongoing
Parkinson's GAD65, GAD67 Intracranial I 12 Complete[19]
Canavan's AAC Intracranial I 21 Ongoing
Batten's CLN2 Intracranial I 10 Ongoing
Alzheimer's NGF Intracranial I 6 Ongoing
Congestive Heart Failure SERCA2a Intra-coronary IIb 250 Ongoing

Trials for the treatment of prostate cancer have reached phase III,[18] however these ex vivo studies do not involve direct administration of AAV to patients.


AAV is not considered to have any known role in disease. It has been suggested to have a role in male infertility,[20] as AAV DNA is more commonly found in semen samples from men with abnormal semen. However, no causal link has been found between AAV infection and male infertility.

AAV structure

AAV genome, transcriptome and proteome

The AAV genome is built of single-stranded deoxyribonucleic acid (ssDNA), either positive- or negative-sensed, which is about 4.7 kilobase long. The genome comprises inverted terminal repeats (ITRs) at both ends of the DNA strand, and two open reading frames (ORFs): rep and cap. The former is composed of four overlapping genes encoding Rep proteins required for the AAV life cycle, and the latter contains overlapping nucleotide sequences of capsid proteins: VP1, VP2 and VP3, which interact together to form a capsid of an icosahedral symmetry.[21]

ITR sequences

The Inverted Terminal Repeat (ITR) sequences comprise 145 bases each. They were named so because of their symmetry, which was shown to be required for efficient multiplication of the AAV genome.[22] The feature of these sequences that gives them this property is their ability to form a hairpin, which contributes to so-called self-priming that allows primase-independent synthesis of the second DNA strand. The ITRs were also shown to be required for both integration of the AAV DNA into the host cell genome (19th chromosome in humans) and rescue from it,[23][24] as well as for efficient encapsidation of the AAV DNA combined with generation of a fully assembled, deoxyribonuclease-resistant AAV particles.[25]

With regard to gene therapy, ITRs seem to be the only sequences required in cis next to the therapeutic gene: structural (cap) and packaging (rep) proteins can be delivered in trans. With this assumption many methods were established for efficient production of recombinant AAV (rAAV) vectors containing a reporter or therapeutic gene. However, it was also published that the ITRs are not the only elements required in cis for the effective replication and encapsidation. A few research groups have identified a sequence designated cis-acting Rep-dependent element (CARE) inside the coding sequence of the rep gene. CARE was shown to augment the replication and encapsidation when present in cis.[26][27][28][29]

rep genes and Rep proteins

On the "left side" of the genome there are two promoters called p5 and p19, from which two overlapping messenger ribonucleic acids (mRNAs) of different length can be produced. Each of these contains an intron which can be either spliced out or not. Given these possibilities, four various mRNAs, and consequently four various Rep proteins with overlapping sequence can be synthesized. Their names depict their sizes in kilodaltons (kDa): Rep78, Rep68, Rep52 and Rep40.[30] Rep78 and 68 can specifically bind the hairpin formed by the ITR in the self-priming act and cleave at a specific region, designated terminal resolution site, within the hairpin. They were also shown to be necessary for the AAVS1-specific integration of the AAV genome. All four Rep proteins were shown to bind ATP and to possess helicase activity. It was also shown that they upregulate the transcription from the p40 promoter (mentioned below), but downregulate both p5 and p19 promoters.[24][30][31][32][33][34]

cap genes and VP proteins

The right side of a positive-sensed AAV genome encodes overlapping sequences of three capsid proteins, VP1, VP2 and VP3, which start from one promoter, designated p40. The molecular weights of these proteins are 87, 72 and 62 kiloDaltons, respectively.[35] All three of them are translated from one mRNA. After this mRNA is synthesized, it can be spliced in two different manners: either a longer or shorter intron can be excised resulting in the formation of two pools of mRNAs: a 2.3 kb- and a 2.6 kb-long mRNA pool. Usually, especially in the presence of adenovirus, the longer intron is preferred, so the 2.3-kb-long mRNA represents the so-called "major splice". In this form the first AUG codon, from which the synthesis of VP1 protein starts, is cut out, resulting in a reduced overall level of VP1 protein synthesis. The first AUG codon that remains in the major splice is the initiation codon for VP3 protein. However, upstream of that codon in the same open reading frame lies an ACG sequence (encoding threonine) which is surrounded by an optimal Kozak context. This contributes to a low level of synthesis of VP2 protein, which is actually VP3 protein with additional N terminal residues, as is VP1.[36][37][38][39]

AAV2 capsid, shown as a ribbon diagram, with the back half hidden for clarity. One fivefold symmetry axis is shown center.

Since the bigger intron is preferred to be spliced out, and since in the major splice the ACG codon is a much weaker translation initiation signal, the ratio at which the AAV structural proteins are synthesized in vivo is about 1:1:20, which is the same as in the mature virus particle.[40] The unique fragment at the N terminus of VP1 protein was shown to possess the phospholipase A2 (PLA2) activity, which is probably required for the releasing of AAV particles from late endosomes.[41] Muralidhar et al. reported that VP2 and VP3 are crucial for correct virion assembly.[38] More recently, however, Warrington et al. showed VP2 to be unnecessary for the complete virus particle formation and an efficient infectivity, and also presented that VP2 can tolerate large insertions in its N terminus, while VP1 can not, probably because of the PLA2 domain presence.[42]

The AAV capsid is composed of 60 capsid protein subunits, VP1, VP2, and VP3, that are arranged in an icosahedral symmetry in a ratio of 1:1:10, with an estimated size of 3.9 MegaDaltons.[43] The crystal structure of the VP3 protein was determined by Xie, Bue, et al..[44]

AAV serotypes, receptors and native tropism

As of 2006 there have been 11 AAV serotypes described, the 11th in 2004.[45] All of the known serotypes can infect cells from multiple diverse tissue types. Tissue specificity is determined by the capsid serotype and pseudotyping of AAV vectors to alter their tropism range will likely be important to their use in therapy.

Serotype 2

Serotype 2 (AAV2) has been the most extensively examined so far.[46][47][48][49][50][51] AAV2 presents natural tropism towards skeletal muscles,[52] neurons,[46] vascular smooth muscle cells[53] and hepatocytes.[54]

Three cell receptors have been described for AAV2: heparan sulfate proteoglycan (HSPG), aVβ5 integrin and fibroblast growth factor receptor 1 (FGFR-1). The first functions as a primary receptor, while the latter two have a co-receptor activity and enable AAV to enter the cell by receptor-mediated endocytosis.[55][56][57] These study results have been disputed by Qiu, Handa, et al..[58] HSPG functions as the primary receptor, though its abundance in the extracellular matrix can scavenge AAV particles and impair the infection efficiency.[59]

Serotype 2 and cancer

Studies have shown that serotype 2 of the virus (AAV-2) apparently kills cancer cells without harming healthy ones. "Our results suggest that adeno-associated virus type 2, which infects the majority of the population but has no known ill effects, kills multiple types of cancer cells yet has no effect on healthy cells," said Craig Meyers,[60] a professor of immunology and microbiology at the Penn State College of Medicine in Pennsylvania.[61] This could lead to a new anti-cancer agent.

Other Serotypes

Although AAV2 is the most popular serotype in various AAV-based research, it has been shown that other serotypes can be more effective as gene delivery vectors. For instance AAV6 appears much better in infecting airway epithelial cells,[62][63] AAV7 presents very high transduction rate of murine skeletal muscle cells (similarly to AAV1 and AAV5), AAV8 is superb in transducing hepatocytes[64][65][66] and AAV1 and 5 were shown to be very efficient in gene delivery to vascular endothelial cells.[67] In the brain, most AAV serotypes show neuronal tropism, while AAV5 also transduces astrocytes.[68] AAV6, a hybrid of AAV1 and AAV2,[66] also shows lower immunogenicity than AAV2.[65]

Serotypes can differ with the respect to the receptors they are bound to. For example AAV4 and AAV5 transduction can be inhibited by soluble sialic acids (of different form for each of these serotypes),[69] and AAV5 was shown to enter cells via the platelet-derived growth factor receptor.[70]

AAV immunology

AAV is of particular interest to gene therapists due to its apparent limited capacity to induce immune responses in humans, a factor which should positively influence vector transduction efficiency while reducing the risk of any immune-associated pathology.


The innate immune response to the AAV vectors has been characterised in animal models. Intravenous administration in mice causes transient production of pro-inflammatory cytokines and some infiltration of neutrophils and other leukocytes into the liver, which seems to sequester a large percentage of the injected viral particles. Both soluble factor levels and cell infiltration appear to return to baseline within six hours. By contrast, more aggressive viruses produce innate responses lasting 24 hours or longer.[71]


The virus is known to instigate robust humoral immunity in animal models and in the human population where up to 80% of individuals are thought to be seropositive for AAV2. Antibodies are known to be neutralising and for gene therapy applications these do impact on vector transduction efficiency via some routes of administration. As well as persistent AAV specific antibody levels, it appears from both prime-boost studies in animals and from clinical trials that the B-cell memory is also strong.[72] In seropositive humans, circulating IgG antibodies for AAV2 appear to be primarily composed of the IgG1 and IgG2 subclasses, with little or no IgG3 or IgG4 present.[73]


The cell-mediated response to the virus and to vectors is poorly characterised and has been largely ignored in the literature as recently as 2005.[72] Clinical trials using an AAV2-based vector to treat haemophilia B seem to indicate that targeted destruction of transduced cells may be occurring.[74] Combined with data that shows that CD8+ T-cells can recognise elements of the AAV capsid in vitro,[75] it appears that there may be a cytotoxic T lymphocyte response to AAV vectors. Cytotoxic responses would imply the involvement of CD4+ T helper cells in the response to AAV and in vitro data from human studies suggests that the virus may indeed induce such responses including both Th1 and Th2 memory responses.[73] A number of candidate T cell stimulating epitopes have been identified within the AAV capsid protein VP1, which may be attractive targets for modification of the capsid if the virus is to be used as a vector for gene therapy.[73][74]

AAV infection cycle

There are several steps in the AAV infection cycle, from infecting a cell to producing new infectious particles:

  1. attachment to the cell membrane
  2. receptor-mediated endocytosis
  3. endosomal trafficking
  4. escape from the late endosome or lysosome
  5. translocation to the nucleus
  6. uncoating
  7. formation of double-stranded DNA replicative form of the AAV genome
  8. expression of rep genes
  9. genome replication
  10. expression of cap genes, synthesis of progeny ssDNA particles
  11. assembly of complete virions, and
  12. release from the infected cell.

Some of these steps may look different in various types of cells, which, in part, contributes to the defined and quite limited native tropism of AAV. Replication of the virus can also vary in one cell type, depending on the cell's current cell cycle phase.[76]

The characteristic feature of the adeno-associated virus is a deficiency in replication and thus its inability to multiply in unaffected cells. The first factor that was described as providing successful generation of new AAV particles, was the adenovirus, from which the AAV name originated. It was then shown that AAV replication can be facilitated by selected proteins derived from the adenovirus genome,[77][78] by other viruses such as HSV,[79] or by genotoxic agents, such as UV irradiation or hydroxyurea.[80][81][82]

The minimal set of the adenoviral genes required for efficient generation of progeny AAV particles, was discovered by Matsushita, Ellinger et al..[77] This discovery allowed for new production methods of recombinant AAV, which do not require adenoviral co-infection of the AAV-producing cells. In the absence of helper virus or genotoxic factors, AAV DNA can either integrate into the host genome or persist in episomal form. In the former case integration is mediated by Rep78 and Rep68 proteins and requires the presence of ITRs flanking the region being integrated. In mice, the AAV genome has been observed persisting for long periods of time in quiescent tissues, such as skeletal muscles, in episomal form (a circular head-to-tail conformation).[83]

See also


  1. ^ Russel DW, Deyle DR (2010). "Adeno- associated virus vector integration". Current Opinion in Molecular Therapy 11 (4): 442–447.  
  2. ^ Grieger JC, Samulski RJ (2005). "Adeno-associated virus as a gene therapy vector: vector development, production and clinical applications". Advances in Biochemical Engineering/biotechnology. Advances in Biochemical Engineering/Biotechnology 99: 119–45.  
  3. ^ Maguire AM, Bennett JB; et al. (2008). "Safety and Efficacy of Gene Transfer for Leber's Congenital Amaurosis". New England Journal of Medicine 358 (21): 2240–8.  
  4. ^ Kotin RM, Siniscalco M, Samulski RJ; et al. (1 March 1990). "Site-specific integration by adeno-associated virus" (PDF). Proc Natl Acad Sci U S A 87 (6): 2211–5.  
  5. ^ Surosky RT, Urabe M, Godwin SG; et al. (1 October 1997). "Adeno-associated virus Rep proteins target DNA sequences to a unique locus in the human genome". Journal of Virology 71 (10): 7951–9.  
  6. ^ Chirmule N, Propert K, Magosin S, Qian Y, Qian R, Wilson J (September 1999). "Immune responses to adenovirus and adeno-associated virus in humans". Gene Therapy 6 (9): 1574–83.  
  7. ^ Hernandez YJ, Wang J, Kearns WG, Loiler S, Poirier A, Flotte TR (1 October 1999). "Latent Adeno-Associated Virus Infection Elicits Humoral but Not Cell-Mediated Immune Responses in a Nonhuman Primate Model". Journal of Virology 73 (10): 8549–58.  
  8. ^ Ponnazhagan S, Mukherjee P, Yoder MC; et al. (April 1997). "Adeno-associated virus 2-mediated gene transfer in vivo: organ-tropism and expression of transduced sequences in mice". Gene 190 (1): 203–10.  
  9. ^ McCarty, D M; Monahan, P E; Samulski, R J (2001). "Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis". Gene Therapy 8 (16): 1248–54.  
  10. ^ Rogers GL (Jan 2014). "Role of the vector genome and underlying factor IX mutation in immune responses to AAV gene therapy for hemophilia B..". J Transl Med. 12.  
  11. ^ J. of Gene Medicine,
  12. ^ Maguire, A. M., Simonelli, F., Pierce, E. A., Pugh, E. N., Mingozzi, F., Bennicelli, J., Banfi, S., et al. (2008). Safety and efficacy of gene transfer for Leber's congenital amaurosis The New England journal of medicine, 358(21), 2240–2248. doi:10.1056/NEJMoa0802315
  13. ^ Bainbridge, J. W. B., Smith, A. J., Barker, S. S., Robbie, S., Henderson, R., Balaggan, K., Viswanathan, A., et al. (2008). Effect of gene therapy on visual function in Leber's congenital amaurosis The New England journal of medicine, 358(21), 2231–2239. doi:10.1056/NEJMoa0802268 PMID 18441371
  14. ^ Hauswirth, W. W., Aleman, T. S., Kaushal, S., Cideciyan, A. V., Schwartz, S. B., Wang, L., Conlon, T. J., et al. (2008). Treatment of Leber Congenital Amaurosis Due to RPE65Mutations by Ocular Subretinal Injection of Adeno-Associated Virus Gene Vector: Short-Term Results of a Phase I Trial. Human gene therapy, 19(10), 979–990. doi:10.1089/hum.2008.107 PMID 18774912
  15. ^ Nathwani, A. C., Tuddenham, E. G. D., Rangarajan, S., Rosales, C., McIntosh, J., Linch, D. C., Chowdary, P., et al. (2011). Adenovirus-associated virus vector-mediated gene transfer in hemophilia B The New England journal of medicine, 365(25), 2357–2365. doi:10.1056/NEJMoa1108046
  16. ^ Jessup, M; Greenberg B, Mancini DM, Cappola T, Pauly DF, Jaski B, Yaroshinsky A, Zsebo K, Dittrich H, Hajjar, RJ (June 2011). "Calcium Upregulation by Percutaneous Administration of Gene Therapy in Cardiac Disease (CUPID): a phase 2 trial of intracoronary gene therapy of sarcoplasmic reticulum Ca2+-ATPase in patients with advanced heart failure". Circulation 124 (3): 304–313.  
  17. ^ LeWitt, P. A., Rezai, A. R., Leehey, M. A., Ojemann, S. G., Flaherty, A. W., Eskandar, E. N., Kostyk, S. K., et al. (2011). AAV2-GAD gene therapy for advanced Parkinson's disease: a double-blind, sham-surgery controlled, randomised trial. The Lancet Neurology, 10(4), 309–319. doi:10.1016/S1474-4422(11)70039-4 PMID 21419704
  18. ^ a b Carter BJ (May 2005). "Adeno-associated virus vectors in clinical trials". Human Gene Therapy 16 (5): 541–50.  
  19. ^ Kaplitt MG, Feigin A, Tang C; et al. (June 2007). "Safety and tolerability of gene therapy with an adeno-associated virus (AAV) borne GAD gene for Parkinson's disease: an open label, phase I trial". Lancet 369 (9579): 2097–105.  
  20. ^ Erles K, Rohde V, Thaele M, Roth S, Edler L, Schlehofer JR (November 2001). "DNA of adeno-associated virus (AAV) in testicular tissue and in abnormal semen samples". Human Reproduction 16 (11): 2333–7.  
  21. ^ Carter, BJ (2000). "Adeno-associated virus and adeno-associated virus vectors for gene delivery". In DD Lassic & N Smyth Templeton. Gene Therapy: Therapeutic Mechanisms and Strategies. New York City: Marcel Dekker, Inc. pp. 41–59.  
  22. ^ Bohenzky RA, LeFebvre RB, Berns KI (October 1988). "Sequence and symmetry requirements within the internal palindromic sequences of the adeno-associated virus terminal repeat". Virology 166 (2): 316–27.  
  23. ^ Wang XS, Ponnazhagan S, Srivastava A (July 1995). "Rescue and replication signals of the adeno-associated virus 2 genome". Journal of Molecular Biology 250 (5): 573–80.  
  24. ^ a b Weitzman MD, Kyöstiö SR, Kotin RM, Owens RA (June 1994). "Adeno-associated virus (AAV) Rep proteins mediate complex formation between AAV DNA and its integration site in human DNA". Proceedings of the National Academy of Sciences of the United States of America 91 (13): 5808–12.  
  25. ^ Zhou X, Muzyczka N (1 April 1998). "In Vitro Packaging of Adeno-Associated Virus DNA". Journal of Virology 72 (4): 3241–7.  
  26. ^ Nony P, Tessier J, Chadeuf G; et al. (October 2001). "Novel cis-Acting Replication Element in the Adeno-Associated Virus Type 2 Genome Is Involved in Amplification of Integrated rep-cap Sequences". Journal of Virology 75 (20): 9991–4.  
  27. ^ Nony P, Chadeuf G, Tessier J, Moullier P, Salvetti A (January 2003). "Evidence for Packaging of rep-cap Sequences into Adeno-Associated Virus (AAV) Type 2 Capsids in the Absence of Inverted Terminal Repeats: a Model for Generation of rep-Positive AAV Particles". Journal of Virology 77 (1): 776–81.  
  28. ^ Philpott NJ, Giraud-Wali C, Dupuis C; et al. (June 2002). "Efficient Integration of Recombinant Adeno-Associated Virus DNA Vectors Requires a p5-rep Sequence in cis". Journal of Virology 76 (11): 5411–21.  
  29. ^ Tullis GE, Shenk T (December 2000). "Efficient Replication of Adeno-Associated Virus Type 2 Vectors: a cis-Acting Element outside of the Terminal Repeats and a Minimal Size". Journal of Virology 74 (24): 11511–21.  
  30. ^ a b Kyöstiö SR, Owens RA, Weitzman MD, Antoni BA, Chejanovsky N, Carter BJ (1 May 1994). "Analysis of adeno-associated virus (AAV) wild-type and mutant Rep proteins for their abilities to negatively regulate AAV p5 and p19 mRNA levels". Journal of Virology 68 (5): 2947–57.  
  31. ^ Im DS, Muzyczka N (May 1990). "The AAV origin binding protein Rep68 is an ATP-dependent site-specific endonuclease with DNA helicase activity". Cell 61 (3): 447–57.  
  32. ^ Im DS, Muzyczka N (1 February 1992). "Partial purification of adeno-associated virus Rep78, Rep52, and Rep40 and their biochemical characterization". Journal of Virology 66 (2): 1119–28.  
  33. ^ Samulski RJ (2003). "AAV vectors, the future workhorse of human gene therapy". Ernst Schering Research Foundation Workshop (43): 25–40.  
  34. ^ Trempe JP, Carter BJ (1 January 1988). "Regulation of adeno-associated virus gene expression in 293 cells: control of mRNA abundance and translation". Journal of Virology 62 (1): 68–74.  
  35. ^ Jay FT, Laughlin CA, Carter BJ (May 1981). "Eukaryotic translational control: adeno-associated virus protein synthesis is affected by a mutation in the adenovirus DNA-binding protein". Proceedings of the National Academy of Sciences of the United States of America 78 (5): 2927–31.  
  36. ^ Becerra SP, Rose JA, Hardy M, Baroudy BM, Anderson CW (December 1985). "Direct mapping of adeno-associated virus capsid proteins B and C: a possible ACG initiation codon". Proceedings of the National Academy of Sciences of the United States of America 82 (23): 7919–23.  
  37. ^ Cassinotti P, Weitz M, Tratschin JD (November 1988). "Organization of the adeno-associated virus (AAV) capsid gene: mapping of a minor spliced mRNA coding for virus capsid protein 1". Virology 167 (1): 176–84.  
  38. ^ a b Muralidhar S, Becerra SP, Rose JA (1 January 1994). "Site-directed mutagenesis of adeno-associated virus type 2 structural protein initiation codons: effects on regulation of synthesis and biological activity". Journal of Virology 68 (1): 170–6.  
  39. ^ Trempe JP, Carter BJ (1 September 1988). "Alternate mRNA splicing is required for synthesis of adeno-associated virus VP1 capsid protein". Journal of Virology 62 (9): 3356–63.  
  40. ^ Rabinowitz JE, Samulski RJ (December 2000). "Building a better vector: the manipulation of AAV virions". Virology 278 (2): 301–8.  
  41. ^ Girod A, Wobus CE, Zádori Z; et al. (1 May 2002). "The VP1 capsid protein of adeno-associated virus type 2 is carrying a phospholipase A2 domain required for virus infectivity". The Journal of General Virology 83 (Pt 5): 973–8.  
  42. ^ Warrington KH, Gorbatyuk OS, Harrison JK, Opie SR, Zolotukhin S, Muzyczka N (June 2004). "Adeno-Associated Virus Type 2 VP2 Capsid Protein Is Nonessential and Can Tolerate Large Peptide Insertions at Its N Terminus". Journal of Virology 78 (12): 6595–609.  
  43. ^ Sonntag F, Schmidt K, Kleinschmidt JA (Jun 2010). "A viral assembly factor promotes AAV2 capsid formation in the nucleolus". Proceedings of the National Academy of Sciences of the United States of America 107 (22): 10220–5.  
  44. ^ Xie Q, Bu W, Bhatia S; et al. (August 2002). "The atomic structure of adeno-associated virus (AAV-2), a vector for human gene therapy". Proceedings of the National Academy of Sciences of the United States of America 99 (16): 10405–10.  
  45. ^ Mori S, Wang L, Takeuchi T, Kanda T (December 2004). "Two novel adeno-associated viruses from cynomolgus monkey: pseudotyping characterization of capsid protein". Virology 330 (2): 375–83.  
  46. ^ a b Bartlett JS, Samulski RJ, McCown TJ (May 1998). "Selective and rapid uptake of adeno-associated virus type 2 in brain". Human Gene Therapy 9 (8): 1181–6.  
  47. ^ Fischer AC, Beck SE, Smith CI; et al. (December 2003). "Successful transgene expression with serial doses of aerosolized rAAV2 vectors in rhesus macaques". Molecular Therapy 8 (6): 918–26.  
  48. ^ Nicklin SA, Buening H, Dishart KL; et al. (September 2001). "Efficient and selective AAV2-mediated gene transfer directed to human vascular endothelial cells". Molecular Therapy 4 (3): 174–81.  
  49. ^ Rabinowitz JE, Xiao W, Samulski RJ (December 1999). "Insertional mutagenesis of AAV2 capsid and the production of recombinant virus". Virology 265 (2): 274–85.  
  50. ^ Shi W, Bartlett JS (April 2003). "RGD inclusion in VP3 provides adeno-associated virus type 2 (AAV2)-based vectors with a heparan sulfate-independent cell entry mechanism". Molecular Therapy 7 (4): 515–25.  
  51. ^ Wu P, Xiao W, Conlon T; et al. (September 2000). "Mutational Analysis of the Adeno-Associated Virus Type 2 (AAV2) Capsid Gene and Construction of AAV2 Vectors with Altered Tropism". Journal of Virology 74 (18): 8635–47.  
  52. ^ Manno CS, Chew AJ, Hutchison S; et al. (April 2003). "AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B". Blood 101 (8): 2963–72.  
  53. ^ Richter M, Iwata A, Nyhuis J; et al. (April 2000). "Adeno-associated virus vector transduction of vascular smooth muscle cells in vivo". Physiological Genomics 2 (3): 117–27.  
  54. ^ Koeberl DD, Alexander IE, Halbert CL, Russell DW, Miller AD (February 1997). "Persistent expression of human clotting factor IX from mouse liver after intravenous injection of adeno-associated virus vectors". Proceedings of the National Academy of Sciences of the United States of America 94 (4): 1426–31.  
  55. ^ Qing K, Mah C, Hansen J, Zhou S, Dwarki V, Srivastava A (January 1999). "Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2". Nature Medicine 5 (1): 71–7.  
  56. ^ Summerford C, Samulski RJ (1 February 1998). "Membrane-Associated Heparan Sulfate Proteoglycan Is a Receptor for Adeno-Associated Virus Type 2 Virions". Journal of Virology 72 (2): 1438–45.  
  57. ^ Summerford C, Bartlett JS, Samulski RJ (January 1999). "AlphaVbeta5 integrin: a co-receptor for adeno-associated virus type 2 infection". Nature Medicine 5 (1): 78–82.  
  58. ^ Qiu J, Handa A, Kirby M, Brown KE (March 2000). "The interaction of heparin sulfate and adeno-associated virus 2". Virology 269 (1): 137–47.  
  59. ^ Pajusola K, Gruchala M, Joch H, Lüscher TF, Ylä-Herttuala S, Büeler H (November 2002). "Cell-Type-Specific Characteristics Modulate the Transduction Efficiency of Adeno-Associated Virus Type 2 and Restrain Infection of Endothelial Cells". Journal of Virology 76 (22): 11530–40.  
  60. ^ "'"Common virus 'kills cancer.  
  61. ^ [2] Science Daily
  62. ^ Limberis, MP; Vandenberghe, LH; Zhang, L; Pickles, RJ; Wilson, JM (February 2009). "Transduction efficiencies of novel AAV vectors in mouse airway epithelium in vivo and human ciliated airway epithelium in vitro.". Molecular therapy : the journal of the American Society of Gene Therapy 17 (2): 294–301.  
  63. ^ Strobel, B; Duechs, MJ; Schmid, R; Stierstorfer, BE; Bucher, H; Quast, K; Stiller, D; Hildebrandt, T; Mennerich, D; Gantner, F; Erb, KJ; Kreuz, S (6 April 2015). "Modeling Pulmonary Disease Pathways Using Recombinant Adeno-associated Virus 6.2.". American journal of respiratory cell and molecular biology.  
  64. ^ Gao GP, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson JM (September 2002). "Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy". Proceedings of the National Academy of Sciences of the United States of America 99 (18): 11854–9.  
  65. ^ a b Halbert CL, Allen JM, Miller AD (July 2001). "Adeno-Associated Virus Type 6 (AAV6) Vectors Mediate Efficient Transduction of Airway Epithelial Cells in Mouse Lungs Compared to That of AAV2 Vectors". Journal of Virology 75 (14): 6615–24.  
  66. ^ a b Rabinowitz JE, Bowles DE, Faust SM, Ledford JG, Cunningham SE, Samulski RJ (May 2004). "Cross-Dressing the Virion: the Transcapsidation of Adeno-Associated Virus Serotypes Functionally Defines Subgroups". Journal of Virology 78 (9): 4421–32.  
  67. ^ Chen S, Kapturczak M, Loiler SA; et al. (February 2005). "Efficient Transduction of Vascular Endothelial Cells with Recombinant Adeno-Associated Virus Serotype 1 and 5 Vectors". Human Gene Therapy 16 (2): 235–47.  
  68. ^ Ortinski P, Dong J, Mungenast A; et al. (May 2010). "Selective induction of astrocytic gliosis generates deficits in neuronal inhibition". Nature Neuroscience 13 (5): 584–91.  
  69. ^ Kaludov N, Brown KE, Walters RW, Zabner J, Chiorini JA (August 2001). "Adeno-Associated Virus Serotype 4 (AAV4) and AAV5 Both Require Sialic Acid Binding for Hemagglutination and Efficient Transduction but Differ in Sialic Acid Linkage Specificity". Journal of Virology 75 (15): 6884–93.  
  70. ^ Di Pasquale G, Davidson BL, Stein CS; et al. (October 2003). "Identification of PDGFR as a receptor for AAV-5 transduction". Nature Medicine 9 (10): 1306–12.  
  71. ^ Zaiss AK, Liu Q, Bowen GP, Wong NC, Bartlett JS, Muruve DA (May 2002). "Differential Activation of Innate Immune Responses by Adenovirus and Adeno-Associated Virus Vectors". Journal of Virology 76 (9): 4580–90.  
  72. ^ a b Zaiss AK, Muruve DA (June 2005). "Immune responses to adeno-associated virus vectors". Current Gene Therapy 5 (3): 323–31.  
  73. ^ a b c Madsen, D.; Cantwell, E.R.; O'Brien, T.; Johnson, P.A.; Mahon, B.P. (2009). "Adeno-associated virus serotype 2 induces cell-mediated immune responses directed against multiple epitopes of the capsid protein VP1". Journal of General Virology 90 (11): 2622–2633.  
  74. ^ a b Manno CS, Pierce GF, Arruda VR; et al. (March 2006). "Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response". Nature Medicine 12 (3): 342–7.  
  75. ^ Sabatino DE, Mingozzi F, Hui DJ; et al. (December 2005). "Identification of mouse AAV capsid-specific CD8+ T cell epitopes". Molecular Therapy 12 (6): 1023–33.  
  76. ^ Rohr UP, Kronenwett R, Grimm D, Kleinschmidt J, Haas R (September 2002). "Primary human cells differ in their susceptibility to rAAV-2-mediated gene transfer and duration of reporter gene expression". Journal of Virological Methods 105 (2): 265–75.  
  77. ^ a b Matsushita T, Elliger S, Elliger C; et al. (July 1998). "Adeno-associated virus vectors can be efficiently produced without helper virus". Gene Therapy 5 (7): 938–45.  
  78. ^ Myers MW, Laughlin CA, Jay FT, Carter BJ (1 July 1980). "Adenovirus helper function for growth of adeno-associated virus: effect of temperature-sensitive mutations in adenovirus early gene region 2". Journal of Virology 35 (1): 65–75.  
  79. ^ Handa H, Carter BJ (25 July 1979). "Adeno-associated virus DNA replication complexes in herpes simplex virus or adenovirus-infected cells". The Journal of Biological Chemistry 254 (14): 6603–10.  
  80. ^ Yalkinoglu AO, Heilbronn R, Bürkle A, Schlehofer JR, zur Hausen H (1 June 1988). "DNA amplification of adeno-associated virus as a response to cellular genotoxic stress". Cancer Research 48 (11): 3123–9.  
  81. ^ Yakobson B, Koch T, Winocour E (1 April 1987). "Replication of adeno-associated virus in synchronized cells without the addition of a helper virus". Journal of Virology 61 (4): 972–81.  
  82. ^ Yakobson B, Hrynko TA, Peak MJ, Winocour E (1 March 1989). "Replication of adeno-associated virus in cells irradiated with UV light at 254 nm". Journal of Virology 63 (3): 1023–30.  
  83. ^ Duan D, Sharma P, Yang J; et al. (1 November 1998). "Circular Intermediates of Recombinant Adeno-Associated Virus Have Defined Structural Characteristics Responsible for Long-Term Episomal Persistence in Muscle Tissue". Journal of Virology 72 (11): 8568–77.  

External links

This article was sourced from Creative Commons Attribution-ShareAlike License; additional terms may apply. World Heritage Encyclopedia content is assembled from numerous content providers, Open Access Publishing, and in compliance with The Fair Access to Science and Technology Research Act (FASTR), Wikimedia Foundation, Inc., Public Library of Science, The Encyclopedia of Life, Open Book Publishers (OBP), PubMed, U.S. National Library of Medicine, National Center for Biotechnology Information, U.S. National Library of Medicine, National Institutes of Health (NIH), U.S. Department of Health & Human Services, and, which sources content from all federal, state, local, tribal, and territorial government publication portals (.gov, .mil, .edu). Funding for and content contributors is made possible from the U.S. Congress, E-Government Act of 2002.
Crowd sourced content that is contributed to World Heritage Encyclopedia is peer reviewed and edited by our editorial staff to ensure quality scholarly research articles.
By using this site, you agree to the Terms of Use and Privacy Policy. World Heritage Encyclopedia™ is a registered trademark of the World Public Library Association, a non-profit organization.

Copyright © World Library Foundation. All rights reserved. eBooks from Project Gutenberg are sponsored by the World Library Foundation,
a 501c(4) Member's Support Non-Profit Organization, and is NOT affiliated with any governmental agency or department.