Adeno-associated virus terminal repeat (TR) mutant generates self-complementary vectors to overcome the rate-limiting step to transduction in vivo
Abstract
An important limitation of recombinant adeno-associated virus (rAAV) vector efficiency is the requirement of hostcell-mediated synthesis of double-stranded DNA from the single-stranded genome. We have bypassed this step in a specialized self-complementary rAAV (scAAV) vector, by utilizing the tendency of AAV to package DNA dimers when the replicating genome is half the length of the wild type (wt). To produce these vectors efficiently, we have deleted the terminal resolution site (trs) from one rAAV TR, preventing the initiation of replication at the mutated end. These constructs generate single-stranded, inverted repeat genomes, with a wt TR at each end, and a mutated TR in the middle. After uncoating, the viral DNA folds through intramolecular base pairing within the mutant TR, which then proceeds through the genome to form a double-stranded molecule. We have used the scAAV to investigate barriers to rAAV transduction in the mouse liver, muscle and brain. In each tissue, scAAV was characterized by faster onset of gene expression and higher transduction efficiency. This study confirms earlier predictions that complementary-strand DNA synthesis is the primary barrier to rAAV-2 transduction. The scAAV is unaffected by this barrier, and provides an extremely efficient vector for gene transfer into many types of cells in vivo.
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The obstacle of ssAAV complementary strand synthesis has been circumvented by the development of self-complementary AAV (scAAV).
“However, this obstacle has been circumvented by the development of self-complementary AAV (scAAV) [10, 11].”
From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [10]
Cites
Ferrari, F. K., Samulski, T., Shenk, T., & Samulski, R. J. (1996) · Journal of Virology · doi:10.1128/jvi.70.5.3227-3234.1996 · Cited by 761 · see citing works
Self-citationDesign and Packaging of Adeno-Associated Virus Gene Targeting Vectors
Hirata, R. K., & Russell, D. W. (2000) · Journal of Virology · doi:10.1128/jvi.74.10.4612-4620.2000 · Cited by 133 · see citing works
Chen, H., McCarty, D., Bruce, A., Suzuki, K., & Suzuki, K. (1998) · Gene Therapy · doi:10.1038/sj.gt.3300547 · Cited by 87 · see citing works
Self-citation
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Adeno-associated virus vector as a platform for gene therapy delivery
Wang, D., Tai, P. W. L., & Gao, G. (2019) · Nature Reviews Drug Discovery · doi:10.1038/s41573-019-0012-9 · Cited by 2,444 · see citing works
Intravascular AAV9 preferentially targets neonatal neurons and adult astrocytes
Foust, K. D., Nurre, E., Montgomery, C. L., Hernandez, A., Chan, C. M., & Kaspar, B. K. (2009) · Nature Biotechnology · doi:10.1038/nbt.1515 · Cited by 1,456 · see citing works
Viral vector platforms within the gene therapy landscape
Bulcha, J. T., Wang, Y., Ma, H., Tai, P. W. L., & Gao, G. (2021) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-021-00487-6 · Cited by 1,404 · see citing works
Engineering adeno-associated virus vectors for gene therapy
Li, C., & Samulski, R. J. (2020) · Nature Reviews Genetics · doi:10.1038/s41576-019-0205-4 · Cited by 1,175 · see citing works
Self-citationImmune responses to AAV vectors: overcoming barriers to successful gene therapy
Mingozzi, F., & High, K. A. (2013) · Blood · doi:10.1182/blood-2013-01-306647 · Cited by 955 · see citing works
Therapeutic in vivo gene transfer for genetic disease using AAV: Progress and challenges
Mingozzi, F., & High, K. A. (2011) · Nature Reviews Genetics · doi:10.1038/nrg2988 · Cited by 893 · see citing works
Foust, K. D., Wang, X., McGovern, V. L., Braun, L., Bevan, A. K., Haidet, A. M., Le, T. T., Morales, P. R., Rich, M. M., Burghes, A. H. M., et al. (2010) · Nature Biotechnology · doi:10.1038/nbt.1610 · Cited by 688 · see citing works
RetractedSelf-complementary AAV vectors; advances and applications
McCarty, D. M. (2008) · Molecular Therapy · doi:10.1038/mt.2008.171 · Cited by 515 · see citing works
Self-citationGray, S. J., Matagne, V., Bachaboina, L., Yadav, S., Ojeda, S. R., & Samulski, R. J. (2011) · Molecular Therapy · doi:10.1038/mt.2011.72 · Cited by 501 · see citing works
Self-citationDuque, S., Joussemet, B., Riviere, C., Marais, T., Dubreil, L., Douar, A.-M., Fyfe, J., Moullier, P., Colle, M.-A., & Barkats, M. (2009) · Molecular Therapy · doi:10.1038/mt.2009.71 · Cited by 511 · see citing works
Gray, S. J., Foti, S. B., Schwartz, J. W., Bachaboina, L., Taylor-Blake, B., Coleman, J., Ehlers, M. D., Zylka, M. J., McCown, T. J., & Samulski, R. J. (2011) · Human Gene Therapy · doi:10.1089/hum.2010.245 · Cited by 420 · see citing works
Self-citationHigh-efficiency transduction of the mouse retina by tyrosine-mutant AAV serotype vectors
Petrs-Silva, H., Dinculescu, A., Li, Q., Min, S.-H., Chiodo, V., Pang, J.-J., Zhong, L., Zolotukhin, S., Srivastava, A., Lewin, A. S., et al. (2009) · Molecular Therapy · doi:10.1038/mt.2008.269 · Cited by 398 · see citing works
Gray, S. J., Nagabhushan Kalburgi, S., McCown, T. J., & Jude Samulski, R. (2013) · Gene Therapy · doi:10.1038/gt.2012.101 · Cited by 377 · see citing works
Overcoming Preexisting Humoral Immunity to AAV Using Capsid Decoys
Mingozzi, F., Anguela, X. M., Pavani, G., Chen, Y., Davidson, R. J., Hui, D. J., Yazicioglu, M., Elkouby, L., Hinderer, C. J., Faella, A., et al. (2013) · Science Translational Medicine · doi:10.1126/scitranslmed.3005795 · Cited by 355 · see citing works
Gene therapy for neurological disorders: progress and prospects
Deverman, B. E., Ravina, B. M., Bankiewicz, K. S., Paul, S. M., & Sah, D. W. Y. (2018) · Nature Reviews Drug Discovery · doi:10.1038/nrd.2018.110 · Cited by 306 · see citing works
Zhong, L., Li, B., Jayandharan, G., Mah, C. S., Govindasamy, L., Agbandje-McKenna, M., Herzog, R. W., Weigel-Van Aken, K. A., Hobbs, J. A., Zolotukhin, S., et al. (2008) · Virology · doi:10.1016/j.virol.2008.08.027 · Cited by 232 · see citing works
AAV hybrid serotypes: improved vectors for gene delivery
Choi, V., McCarty, D., & Samulski, R. (2005) · Current Gene Therapy · doi:10.2174/1566523054064968 · Cited by 191 · see citing works
Self-citationWu, Z., Sun, J., Zhang, T., Yin, C., Yin, F., Van Dyke, T., Samulski, R. J., & Monahan, P. E. (2008) · Molecular Therapy · doi:10.1038/sj.mt.6300355 · Cited by 153 · see citing works
Self-citationResolving Adeno-Associated Viral Particle Diversity With Charge Detection Mass Spectrometry
Pierson, E. E., Keifer, D. Z., Asokan, A., & Jarrold, M. F. (2016) · Analytical Chemistry · doi:10.1021/acs.analchem.6b00883 · Cited by 152 · see citing works
Allay, J. A., Sleep, S., Long, S., Tillman, D. M., Clark, R., Carney, G., Fagone, P., McIntosh, J. H., Nienhuis, A. W., Davidoff, A. M., et al. (2011) · Human Gene Therapy · doi:10.1089/hum.2010.202 · Cited by 152 · see citing works
Mutation-independent rhodopsin gene therapy by knockdown and replacement with a single AAV vector
Cideciyan, A. V., Sudharsan, R., Dufour, V. L., Massengill, M. T., Iwabe, S., Swider, M., Lisi, B., Sumaroka, A., Marinho, L. F., Appelbaum, T., et al. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1805055115 · Cited by 147 · see citing works
Davidsson, M., Wang, G., Aldrin-Kirk, P., Cardoso, T., Nolbrant, S., Hartnor, M., Mudannayake, J., Parmar, M., & Björklund, T. (2019) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1910061116 · Cited by 142 · see citing works
Mingozzi, F., Chen, Y., Edmonson, S. C., Zhou, S., Thurlings, R. M., Tak, P. P., High, K. A., & Vervoordeldonk, M. J. (2013) · Gene Therapy · doi:10.1038/gt.2012.55 · Cited by 138 · see citing works
AAV vectors applied to the treatment of CNS disorders: Clinical status and challenges
Kang, L., Jin, S., Wang, J., Lv, Z., Xin, C., Tan, C., Zhao, M., Wang, L., & Liu, J. (2023) · Journal of Controlled Release · doi:10.1016/j.jconrel.2023.01.067 · Cited by 131 · see citing works
Ferrari, S., Jacob, A., Cesana, D., Laugel, M., Beretta, S., Varesi, A., Unali, G., Conti, A., Canarutto, D., Albano, L., et al. (2022) · Cell stem cell · doi:10.1016/j.stem.2022.09.001 · Cited by 133 · see citing works
Characterization of a recombinant adeno-associated virus type 2 Reference Standard Material
Lock, M., McGorray, S., Auricchio, A., Ayuso, E., Beecham, E. J., Blouin-Tavel, V., Bosch, F., Bose, M., Byrne, B. J., Caton, T., et al. (2010) · Human Gene Therapy · doi:10.1089/hum.2009.223 · Cited by 131 · see citing works
Self-citationShort DNA Hairpins Compromise Recombinant Adeno-Associated Virus Genome Homogeneity
Xie, J., Mao, Q., Tai, P. W. L., He, R., Ai, J., Su, Q., Zhu, Y., Ma, H., Li, J., Gong, S., et al. (2017) · Molecular Therapy · doi:10.1016/j.ymthe.2017.03.028 · Cited by 102 · see citing works
Methods for Gene Transfer to the Central Nervous System
Kantor, B., Bailey, R. M., Wimberly, K., Kalburgi, S. N., & Gray, S. J. (2014) · Advances in Genetics · doi:10.1016/b978-0-12-800149-3.00003-2 · Cited by 109 · see citing works
Pañeda, A., Vanrell, L., Mauleon, I., Crettaz, J. S., Berraondo, P., Timmermans, E. J., Beattie, S. G., Twisk, J., van Deventer, S., Prieto, J., et al. (2009) · Human Gene Therapy · doi:10.1089/hum.2009.031 · Cited by 109 · see citing works
Tai, P. W. L., Xie, J., Fong, K., Seetin, M., Heiner, C., Su, Q., Weiand, M., Wilmot, D., Zapp, M. L., & Gao, G. (2018) · Molecular Therapy - Methods & Clinical Development · doi:10.1016/j.omtm.2018.02.002 · Cited by 102 · see citing works
Fagone, P., Wright, J. F., Nathwani, A. C., Nienhuis, A. W., Davidoff, A. M., & Gray, J. T. (2012) · Human Gene Therapy Methods · doi:10.1089/hgtb.2011.104 · Cited by 97 · see citing works
Bailey, R. M., Rozenberg, A., & Gray, S. J. (2020) · Brain Research · doi:10.1016/j.brainres.2020.146832 · Cited by 83 · see citing works
Gene therapy for retinal disease
McClements, M. E., & MacLaren, R. E. (2013) · Translational Research · doi:10.1016/j.trsl.2012.12.007 · Cited by 80 · see citing works
Liver induced transgene tolerance with AAV vectors
Keeler, G. D., Markusic, D. M., & Hoffman, B. E. (2019) · Cellular Immunology · doi:10.1016/j.cellimm.2017.12.002 · Cited by 81 · see citing works
Lovric, J., Mano, M., Zentilin, L., Eulalio, A., Zacchigna, S., & Giacca, M. (2012) · Molecular Therapy · doi:10.1038/mt.2012.144 · Cited by 77 · see citing works
Enhancing transduction of the liver by adeno-associated viral vectors
Nathwani, A. C., Cochrane, M., McIntosh, J., Ng, C. Y. C., Zhou, J., Gray, J. T., & Davidoff, A. M. (2009) · Gene Therapy · doi:10.1038/gt.2008.137 · Cited by 77 · see citing works
Pozsgai, E. R., Griffin, D. A., Heller, K. N., Mendell, J. R., & Rodino-Klapac, L. R. (2017) · Molecular Therapy · doi:10.1016/j.ymthe.2017.02.013 · Cited by 76 · see citing works
Tran, N. T., Lecomte, E., Saleun, S., Namkung, S., Robin, C., Weber, K., Devine, E., Blouin, V., Adjali, O., Ayuso, E., et al. (2022) · Human Gene Therapy · doi:10.1089/hum.2022.050 · Cited by 74 · see citing works
Clinical Applications Involving CNS Gene Transfer
Kantor, B., McCown, T., Leone, P., & Gray, S. J. (2014) · Advances in Genetics · doi:10.1016/b978-0-12-800149-3.00002-0 · Cited by 74 · see citing works
Yang, Y., Wu, X.-Q., Li, W.-X., Huang, H.-M., Li, H.-D., Pan, X.-Y., Li, X.-F., Huang, C., Meng, X.-M., Zhang, L., et al. (2018) · Oncogene · doi:10.1038/s41388-018-0383-0 · Cited by 72 · see citing works
Koilkonda, R., Yu, H., Talla, V., Porciatti, V., Feuer, W. J., Hauswirth, W. W., Chiodo, V., Erger, K. E., Boye, S. L., Lewin, A. S., et al. (2014) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.14-15388 · Cited by 65 · see citing works
AAV vector-mediated in vivo reprogramming into pluripotency
Senís, E., Mosteiro, L., Wilkening, S., Wiedtke, E., Nowrouzi, A., Afzal, S., Fronza, R., Landerer, H., Abad, M., Niopek, D., et al. (2018) · Nature Communications · doi:10.1038/s41467-018-05059-x · Cited by 64 · see citing works
Production of recombinant adeno-associated viral vectors for in vitro and in vivo use
Choi, V. W., Asokan, A., Haberman, R. A., & Samulski, R. J. (2007) · Current Protocols in Molecular Biology · doi:10.1002/0471142727.mb1625s78 · Cited by 52 · see citing works
Self-citationRational engineering of a functional CpG-free ITR for AAV gene therapy
Pan, X., Yue, Y., Boftsi, M., Wasala, L. P., Tran, N. T., Zhang, K., Pintel, D. J., Tai, P. W. L., & Duan, D. (2022) · Gene Therapy · doi:10.1038/s41434-021-00296-0 · Cited by 53 · see citing works
Karumuthil‐Melethil, S., Marshall, M. S., Heindel, C., Jakubauskas, B., Bongarzone, E. R., & Gray, S. J. (2016) · Journal of Neuroscience Research · doi:10.1002/jnr.23882 · Cited by 51 · see citing works
Koilkonda, R. D. (2010) · Archives of Ophthalmology · doi:10.1001/archophthalmol.2010.135 · Cited by 48 · see citing works
Xu, L., Yao, S., Ding, Y. E., Xie, M., Feng, D., Sha, P., Tan, L., Bei, F., & Yao, Y. (2024) · Journal of Translational Medicine · doi:10.1186/s12967-024-05661-2 · Cited by 48 · see citing works
Rogers, G. L., Martino, A. T., Zolotukhin, I., Ertl, H. C., & Herzog, R. W. (2014) · Journal of Translational Medicine · doi:10.1186/1479-5876-12-25 · Cited by 45 · see citing works
Mitchell, A. M., Li, C., & Samulski, R. J. (2013) · Journal of Virology · doi:10.1128/jvi.03443-12 · Cited by 41 · see citing works
Self-citationFrancis, J. S., Markov, V., Wojtas, I. D., Gray, S., McCown, T., Samulski, R. J., Figueroa, M., & Leone, P. (2021) · Molecular Therapy - Methods & Clinical Development · doi:10.1016/j.omtm.2021.01.009 · Cited by 41 · see citing works
Self-citationAAV-mediated RLBP1 gene therapy improves the rate of dark adaptation in Rlbp1 knockout mice
Choi, V. W., Bigelow, C. E., McGee, T. L., Gujar, A. N., Li, H., Hanks, S. M., Vrouvlianis, J., Maker, M., Leehy, B., Zhang, Y., et al. (2015) · Molecular Therapy - Methods & Clinical Development · doi:10.1038/mtm.2015.22 · Cited by 35 · see citing works
Bell, P., Wang, L., Chen, S.-J., Yu, H., Zhu, Y., Nayal, M., He, Z., White, J., Lebel-Hagan, D., & Wilson, J. M. (2016) · Human Gene Therapy Methods · doi:10.1089/hgtb.2016.039 · Cited by 26 · see citing works
Fagoe, N. D., Eggers, R., Verhaagen, J., & Mason, M. R. J. (2014) · Gene Therapy · doi:10.1038/gt.2013.71 · Cited by 27 · see citing works
Our journey to successful gene therapy for hemophilia B
Nathwani, A. C., Nienhuis, A. W., & Davidoff, A. M. (2014) · Human Gene Therapy · doi:10.1089/hum.2014.2540 · Cited by 15 · see citing works
- Authors
- McCarty, D. M., Fu, H., Monahan, P. E., Toulson, C. E., Naik, P., & Samulski, R. J.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- Venue
- Gene Therapy
- Published
- 2003-11-17
- Volume
- 10
- Issue
- 26
- Pages
- 2112-2118
- ISSN
- 0969-7128 (ISSN-L)
- 1476-5462
- OpenAlex
- W2081064825
- Cited by
- 547
- References
- 3
- Citations by year
- 2026: 18 · 2025: 14 · 2024: 24 · 2023: 25 · 2022: 23 · 2021: 23 · 2020: 23 · 2019: 22 · 2018: 24 · 2017: 20 · 2016: 21 · 2015: 25 · 2014: 28 · 2013: 36 · 2012: 33
- FWCI
- 6.36
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Heart, Lung, and Blood Institute (P01HL051818, HL 03960, P01-HL51818-06, P01HL066973, P01-HL66973-01)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- adeno-associated virus, terminal repeats, self-complementary AAV vectors, intramolecular base pairing, transduction efficiency, in vivo gene delivery, liver, muscle, brain
- MeSH terms
- Dependovirus, Animals, Brain, DNA Replication, DNA, Viral, Genetic Vectors, Transduction, Genetic, Gene Deletion, Terminal Repeat Sequences, Hepatocytes, Muscle Cells, Mice
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