Hybrid adeno-associated virus bearing nonhomologous inverted terminal repeats enhances dual-vector reconstruction of minigenes in vivo

Ziying Yan; Diana C M Lei-Butters; Yulong Zhang; Roman Zak; John F Engelhardt

doi:10.1089/hum.2006.128

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Hybrid adeno-associated virus bearing nonhomologous inverted terminal repeats enhances dual-vector reconstruction of minigenes in vivo

Journal article

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Peer reviewed

Hybrid adeno-associated virus bearing nonhomologous inverted terminal repeats enhances dual-vector reconstruction of minigenes in vivo

Ziying Yan, Diana C M Lei-Butters, Yulong Zhang, Roman Zak and John F Engelhardt

Human gene therapy, Vol.18(1), pp.81-87

01/2007

DOI: 10.1089/hum.2006.128

PMCID: PMC2121583

PMID: 17181493

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https://www.ncbi.nlm.nih.gov/pmc/articles/2121583View

Open Access

Abstract

We have previously demonstrated that hybrid adeno-associated viral (AAV) vectors bearing nonhomologous inverted terminal repeats (ITRs) enhance directional intermolecular recombination and the efficiency of dual-AAV vector trans-splicing in cultured cells. Using hybrid-ITR vectors carrying two exons of a lacZ minigene, we demonstrate that this dual-vector approach also mediates higher levels (3- to 6-fold) of gene reconstitution in mouse skeletal muscle, liver, and heart. Inhibition of the proteasome by systemic administration of Doxil (Food and Drug Administration-approved lipid-formulated doxorubicin) further enhanced dual-vector trans-splicing 6- to 12-fold in two mouse strains. Hence, using hybrid-ITR AAV vectors in combination with proteasome modulation enhanced dual-vector delivery of a transgene approximately 36-fold over the current dual-vector trans-splicing approaches. These data provide in vivo evidence that ITR sequence-dependent homologous recombination, rather than nonhomologous end joining, is the predominant mechanism for AAV genome heterodimerization. Hence, enhanced directional recombination provided by hybrid-ITR vectors may be a useful in vivo strategy for improving dual-vector delivery of transgenes larger than the AAV packaging limit.

Cell Line

Gene Expression Regulation, Viral - drug effects

Transduction, Genetic

Antibiotics, Antineoplastic - pharmacology

Genome, Viral - genetics

Gene Expression Regulation, Viral - genetics

Dependovirus

Trans-Splicing - drug effects

Animals

Mice

Genetic Vectors

Terminal Repeat Sequences

Transgenes

Doxorubicin - pharmacology

Lac Operon

Details

Title: Subtitle: Hybrid adeno-associated virus bearing nonhomologous inverted terminal repeats enhances dual-vector reconstruction of minigenes in vivo
Creators: Ziying Yan - Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242, USA
Diana C M Lei-Butters
Yulong Zhang
Roman Zak
John F Engelhardt
Resource Type: Journal article
Publication Details: Human gene therapy, Vol.18(1), pp.81-87
DOI: 10.1089/hum.2006.128
PMID: 17181493
PMCID: PMC2121583
NLM abbreviation: Hum Gene Ther
ISSN: 1043-0342
eISSN: 1557-7422
Publisher: United States
Grant note: R01 HL058340 / NHLBI NIH HHS R01 HL058340-11 / NHLBI NIH HHS P30 DK054759 / NIDDK NIH HHS R01 HL58340 / NHLBI NIH HHS DK54759 / NIDDK NIH HHS
Language: English
Date published: 01/2007
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
Record Identifier: 9984025302302771

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