Viral Hybrid Vectors for Somatic Integration - Are They the Better Solution?

Müther, Nadine; Noske, Nadja; Ehrhardt, Anja

doi:10.3390/v1031295

Cited by 15 publications

(13 citation statements)

References 143 publications

(181 reference statements)

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“…In contrast to other previously described adenoviral hybrid-vector systems, 3,37 our vector releases a genetic element that can be maintained episomally after excision based on FLPe recombination. In this study, we demonstrated that the plasmid replicon pEPito can function as a substrate for FLPe recombination and, at the same time, maintain its function as a plasmid replicon for stable transgene expression.…”

Section: Discussionmentioning

confidence: 99%

A Novel Adenoviral Hybrid-vector System Carrying a Plasmid Replicon for Safe and Efficient Cell and Gene Therapeutic Applications

Voigtlander

Haase

Mück-Häusl

et al. 2013

Molecular Therapy - Nucleic Acids

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In dividing cells, the two aims a gene therapeutic approach should accomplish are efficient nuclear delivery and retention of therapeutic DNA. For stable transgene expression, therapeutic DNA can either be maintained by somatic integration or episomal persistence of which the latter approach would diminish the risk of insertional mutagenesis. As most monosystems fail to fulfill both tasks with equal efficiency, hybrid-vector systems represent promising alternatives. Our hybrid-vector system synergizes high-capacity adenoviral vectors (HCAdV) for efficient delivery and the scaffold/matrix attachment region (S/MAR)–based pEPito plasmid replicon for episomal persistence. After proving that this plasmid replicon can be excised from adenovirus in vitro, colony forming assays were performed. We found an increased number of colonies of up to sevenfold in cells that received the functional plasmid replicon proving that the hybrid-vector system is functional. Transgene expression could be maintained for 6 weeks and the extrachromosomal plasmid replicon was rescued. To show efficacy in vivo, the adenoviral hybrid-vector system was injected into C57Bl/6 mice. We found that the plasmid replicon can be released from adenoviral DNA in murine liver resulting in long-term transgene expression. In conclusion, we demonstrate the efficacy of our novel HCAdV-pEPito hybrid-vector system in vitro and in vivo.

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Section: Discussionmentioning

confidence: 99%

A Novel Adenoviral Hybrid-vector System Carrying a Plasmid Replicon for Safe and Efficient Cell and Gene Therapeutic Applications

Voigtlander

Haase

Mück-Häusl

et al. 2013

Molecular Therapy - Nucleic Acids

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“…Since that time, several studies have been developed on hybrid transposase systems (reviewed in Refs. [60,61]) that use adenovirus [62][63][64], adeno-associated virus [65], baculovirus [66], or nonintegrative lentivirus [67,68] particles.…”

Section: Viral Hybrid Systemsmentioning

confidence: 99%

DNA Elements Tetris: A Strategy for Gene Correction

Bastie¹,

Rouleux‐Bonnin²

2016

Modern Tools for Genetic Engineering

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Transposable elements (TEs) are mobile genetic sequences that are able to move in the genome from one location to another. TEs were first regarded as junk or selfish DNA, as they comprise the largest molecular class within most metazoan genomes having no genomic function. It was necessary to wait until whole genome sequencing to provide new insights about the origin, diversity, and impact of TEs on the genome function. Thus, due to advances in molecular technology, TEs have been shown to create new regulatory sequence networks. Although nowadays most TEs present in the human genome are silenced, particularly DNA transposons, it does not mean that these sequences are dead. In this review, we detail how DNA transposons could be emphasized to create a new tool for gene correction. DNA-based transposon vectors are derived from three models: Sleeping Beauty, piggyBac, and Tol2, which all work via a "cut-and-paste" mechanism where transposase enzyme is alone able to catalyze the transposition process, which means integrating the genes of interest in chromosomal DNA. Limitations and improvements of the systems are discussed, particularly the latest way to target a specific integration site, showing that the DNA transposon-derived system and its engineering, are powerful tools for gene correction.

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“…Hybrid Ad systems have been described with elements incorporated from adeno-associated virus (AAV), retrovirus, sleeping beauty (SB) transposons, L1 retrotransposons, or PhiC31 integrase. 4 While each of these strategies successfully facilitates genomic integration of cargo genes, efficiencies are typically below 1% of transduced cells. [5][6][7][8] A hybrid Ad-lentiviral system has achieved an in vivo stable transduction rate of 20-30% in mouse liver, 9 although the efficiency was likely influenced by the immunodeficient nature of the mice that were used.…”

Section: Introductionmentioning

confidence: 99%

A Hybrid Adenoviral Vector System Achieves Efficient Long-Term Gene Expression in the Liver viapiggyBacTransposition

et al. 2015

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Much research has gone into the development of hybrid gene delivery systems that combine the broad tropism and efficient transduction of adenoviral vectors with the ability to achieve stable expression of cargo genes. In addition to gene therapy applications, such a system has considerable advantages for studies of gene function in vivo, permitting fine-tuned genetic manipulation with higher throughput than can be achieved using standard transgenic and DNA targeting techniques. Existing strategies are limited, however, by low integration efficiencies, small cargo capacity, and/or a dependence on target cell division. The utility of this approach could be enhanced by a system that provides all of the following: (1) efficient delivery, (2) stable expression in a high percentage of target cells (whether mitotic or not), (3) large cargo capacity, (4) flexibility to use with a wide range of additional experimental conditions, and (5) simple experimental technique. Here we report the initial characterization of a hybrid system that meets these criteria by utilizing piggyBac (PB) transposition to achieve genomic integration from adenoviral vectors. We demonstrate stable expression of an adenovirus (Ad)-PBdelivered reporter gene in *20-40% of hepatocytes following standard tail vein injection. Its high efficiency and flexibility relative to existing hybrid adenoviral gene delivery approaches indicate a considerable potential utility of the Ad-PB system for therapeutic gene delivery and in vivo studies of gene function.

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Viral Hybrid Vectors for Somatic Integration - Are They the Better Solution?

Cited by 15 publications

References 143 publications

A Novel Adenoviral Hybrid-vector System Carrying a Plasmid Replicon for Safe and Efficient Cell and Gene Therapeutic Applications

A Novel Adenoviral Hybrid-vector System Carrying a Plasmid Replicon for Safe and Efficient Cell and Gene Therapeutic Applications

DNA Elements Tetris: A Strategy for Gene Correction

A Hybrid Adenoviral Vector System Achieves Efficient Long-Term Gene Expression in the Liver viapiggyBacTransposition

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