TraFo-CRISPR: Enhanced Genome Engineering by Transient Foamy Virus Vector-Mediated Delivery of CRISPR/Cas9 Components
The adaptation of CRISPR/Cas technology for use in mammals has revolutionized genome engineering. In particular with regard to clinical application, efficient expression of Cas9 within a narrow time frame is highly desirable to minimize the accumulation of off-target editing. We developed an effecti...
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| Main Authors: | , , , , , , , , , , |
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| Format: | Book |
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Elsevier,
2019-12-01T00:00:00Z.
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|---|---|---|---|
| 001 | doaj_36bae1567ab84d818e9e97c53017f3b3 | ||
| 042 | |a dc | ||
| 100 | 1 | 0 | |a Fabian Lindel |e author |
| 700 | 1 | 0 | |a Carolin R. Dodt |e author |
| 700 | 1 | 0 | |a Niklas Weidner |e author |
| 700 | 1 | 0 | |a Monique Noll |e author |
| 700 | 1 | 0 | |a Fabian Bergemann |e author |
| 700 | 1 | 0 | |a Rayk Behrendt |e author |
| 700 | 1 | 0 | |a Sarah Fischer |e author |
| 700 | 1 | 0 | |a Josephine Dietrich |e author |
| 700 | 1 | 0 | |a Marc Cartellieri |e author |
| 700 | 1 | 0 | |a Martin V. Hamann |e author |
| 700 | 1 | 0 | |a Dirk Lindemann |e author |
| 245 | 0 | 0 | |a TraFo-CRISPR: Enhanced Genome Engineering by Transient Foamy Virus Vector-Mediated Delivery of CRISPR/Cas9 Components |
| 260 | |b Elsevier, |c 2019-12-01T00:00:00Z. | ||
| 500 | |a 2162-2531 | ||
| 500 | |a 10.1016/j.omtn.2019.10.006 | ||
| 520 | |a The adaptation of CRISPR/Cas technology for use in mammals has revolutionized genome engineering. In particular with regard to clinical application, efficient expression of Cas9 within a narrow time frame is highly desirable to minimize the accumulation of off-target editing. We developed an effective, aptamer-independent retroviral delivery system for Cas9 mRNAs that takes advantage of a unique foamy virus (FV) capability: the efficient encapsidation and transfer of non-viral RNAs. This enabled us to create a FV vector toolbox for efficient, transient delivery (TraFo) of CRISPR/Cas9 components into different target tissues. Co-delivery of Cas9 mRNA by TraFo-Cas9 vectors in combination with retroviral, integration-deficient single guide RNA (sgRNA) expression enhanced efficacy and specificity of gene-inactivation compared with CRISPR/Cas9 lentiviral vector systems. Furthermore, separate TraFo-Cas9 delivery allowed the optional inclusion of a repair matrix for efficient gene correction or tagging as well as the addition of fluorescent negative selection markers for easy identification of off-target editing or incorrect repair events. Thus, the TraFo CRISPR toolbox represents an interesting alternative technology for gene inactivation and gene editing. Keywords: CRISPR/Cas9, genome engineering, viral vector, transient system, TraFo-Cas9, foamy virus, CAR T-cell | ||
| 546 | |a EN | ||
| 690 | |a Therapeutics. Pharmacology | ||
| 690 | |a RM1-950 | ||
| 655 | 7 | |a article |2 local | |
| 786 | 0 | |n Molecular Therapy: Nucleic Acids, Vol 18, Iss , Pp 708-726 (2019) | |
| 787 | 0 | |n http://www.sciencedirect.com/science/article/pii/S2162253119303026 | |
| 787 | 0 | |n https://doaj.org/toc/2162-2531 | |
| 856 | 4 | 1 | |u https://doaj.org/article/36bae1567ab84d818e9e97c53017f3b3 |z Connect to this object online. |