Specifically integrating vectors for targeted gene delivery: progress and prospects

Kovač, Adrian; Ivics, Zoltán

doi:10.18609/cgti.2017.013

Cited by 12 publications

(20 citation statements)

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“…Importantly, no SB-associated adverse effects have been observed in preclinical animal studies [26,28,86,87]. Finally, the future development of molecular strategies aiming at targetselected transgene integration with transposon-based vectors [88] may further enhance the safety profile of SB (see Outstanding Questions).…”

Section: Glossarymentioning

confidence: 99%

Gene Therapy with the Sleeping Beauty Transposon System

et al. 2017

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Section: Glossarymentioning

confidence: 99%

Gene Therapy with the Sleeping Beauty Transposon System

et al. 2017

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“…6,8,67,68 Finally, SB's safety profile can probably be further improved through molecular strategies that enhance target-selected transgene integration. 69 …”

Section: Safety Aspects Of Sb Transpositionmentioning

confidence: 99%

Wide Awake and Ready to Move: 20 Years of Non-Viral Therapeutic Genome Engineering with the Sleeping Beauty Transposon System

et al. 2017

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Gene therapies will only become a widespread tool in the clinical treatment of human diseases with the advent of gene transfer vectors that integrate genetic information stably, safely, effectively, and economically. Two decades after the discovery of the Sleeping Beauty (SB) transposon, it has been transformed into a vector system that is fulfilling these requirements. SB may well overcome some of the limitations associated with viral gene transfer vectors and transient non-viral gene delivery approaches that are being used in the majority of ongoing clinical trials. The SB system has achieved a high level of stable gene transfer and sustained transgene expression in multiple primary human somatic cell types, representing crucial steps that may permit its clinical use in the near future. This article reviews the most important aspects of SB as a tool for gene therapy, including aspects of its vectorization and genomic integration. As an illustration, the clinical development of the SB system toward gene therapy of age-related macular degeneration and cancer immunotherapy is highlighted.

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“…The main drawback of integrating vectors is their unspecific or semi-random integration (14). For example, lentiviruses or γ-retroviruses actively target genes or transcriptional start sites (15)(16)(17)(18)(19).…”

Section: Introductionmentioning

confidence: 99%

“…Indeed, by using DBDs to tether integrating enzymes (retroviral integrases or transposases) to the desired target, one can combine the efficient, DSB-free insertion of genetic cargo with the target specificity of designer nucleases [reviewed in (14)]. In general, two approaches can be used to direct transposon integrations by using a DBD: direct fusions or adapter proteins (14). In the direct fusion approach, a fusion protein of a DBD and the transposase is generated to tether the transposase to the target site ( Fig.…”

Section: Introductionmentioning

confidence: 99%

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RNA-guided Retargeting of Sleeping Beauty Transposition in Human Cells

Kovač

Miskey

Menzel³

et al. 2019

Preprint

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Two different approaches of genomic modification are currently used for genome engineering and gene therapy: integrating vectors, which can efficiently integrate large transgenes but are unspecific with respect to their integration sites, and nuclease-based approaches, which are highly specific but not efficient at integrating large genetic cargoes. Here we demonstrate biased genome-wide integration of the Sleeping Beauty (SB) transposon by combining it with components of the CRISPR/Cas9 system. We provide proof-of-concept that it is possible to influence the target site selection of SB by fusing it to a catalytically inactive Cas9 (dCas9) and by providing a single guide RNA (sgRNA) against the human Alu retrotransposon. Enrichment of transposon integrations was dependent on the sgRNA, occurred in a relatively narrow, ~200 bp window around the targeted sites and displayed an asymmetric pattern with a bias towards sites that are downstream of the sgRNA targets. Our data indicate that the targeting mechanism specified by CRISPR/Cas9 forces integration into genomic regions that are otherwise poor targets for SB transposition. Future modifications of this technology may allow the development of methods for efficient and specific gene insertion for precision genetic engineering.

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Specifically integrating vectors for targeted gene delivery: progress and prospects

Cited by 12 publications

References 0 publications

Gene Therapy with the Sleeping Beauty Transposon System

Gene Therapy with the Sleeping Beauty Transposon System

Wide Awake and Ready to Move: 20 Years of Non-Viral Therapeutic Genome Engineering with the Sleeping Beauty Transposon System

RNA-guided Retargeting of Sleeping Beauty Transposition in Human Cells

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