ArticleScientific reports2014
Non-integrating gamma-retroviral vectors as a versatile tool for transient zinc-finger nuclease delivery.
Article in Scientific reports, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 28 citations in OpenAlex.
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- Immune Cell Hacking: Challenges and Clinical Approaches to Create Smarter Generations of Chimeric Antigen Receptor T Cells.Frontiers in immunology · 2018Review
- The Functionality of Minimal PiggyBac Transposons in Mammalian Cells.Molecular therapy. Nucleic acids · 2016Article
- Efficient delivery of nuclease proteins for genome editing in human stem cells and primary cells.Nature protocols · 2015Article
- Improved cell-penetrating zinc-finger nuclease proteins for precision genome engineering.Molecular therapy. Nucleic acids · 2015Article
- Novel lentiviral vectors with mutated reverse transcriptase for mRNA delivery of TALE nucleases.Scientific reports · 2014Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Designer nucleases, like zinc-finger nucleases (ZFNs), represent valuable tools for targeted genome editing. Here, we took advantage of the gamma-retroviral life cycle and produced vectors to transfer ZFNs in the form of protein, mRNA and episomal DNA. Transfer efficacy and ZFN activity were assessed in quantitative proof-of-concept experiments in a human cell line and in mouse embryonic stem cells. We demonstrate that retrovirus-mediated protein transfer (RPT), retrovirus-mediated mRNA transfer (RMT), and retrovirus-mediated episome transfer (RET) represent powerful methodologies for transient protein delivery or protein expression. Furthermore, we describe complementary strategies to augment ZFN activity after gamma-retroviral transduction, including serial transduction, proteasome inhibition, and hypothermia. Depending on vector dose and target cell type, gene disruption frequencies of up to 15% were achieved with RPT and RMT, and >50% gene knockout after RET. In summary, non-integrating gamma-retroviral vectors represent a versatile tool to transiently deliver ZFNs to human and mouse cells.
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