Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles
Abstract Prime Editing can rewrite genes in living cells by allowing point mutations, deletions, or insertion of small DNA sequences with high precision. However, its safe and efficient delivery into human stem cells remains a technical challenge. In this report, we engineer Nanoscribes, virus-like...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55604-0 |
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| author | Thibaut Halegua Valérie Risson Julien Carras Martin Rouyer Laurent Coudert Arnaud Jacquier Laurent Schaeffer Théophile Ohlmann Philippe Emmanuel Mangeot |
| author_facet | Thibaut Halegua Valérie Risson Julien Carras Martin Rouyer Laurent Coudert Arnaud Jacquier Laurent Schaeffer Théophile Ohlmann Philippe Emmanuel Mangeot |
| author_sort | Thibaut Halegua |
| collection | DOAJ |
| description | Abstract Prime Editing can rewrite genes in living cells by allowing point mutations, deletions, or insertion of small DNA sequences with high precision. However, its safe and efficient delivery into human stem cells remains a technical challenge. In this report, we engineer Nanoscribes, virus-like particles that encapsidate ribonucleoprotein complexes of the Prime Editing system and allow their delivery into recipient cells. We identify key features that unlock the potential of Nanoscribes, including the use of multiple fusogens, the improvement of pegRNAs structures, their encoding by a Pol II system and the optimization of Prime-Editors. Nanoscribes edit HEK293T with an efficiency of 68% at the HEK3 locus with increased fidelity over DNA-transfection and support pegRNA-multiplexing. Importantly, Nanoscribes permit editing of myoblasts, hiPSCs and hiPSCs-derived hematopoietic stem cells with an editing efficiency up to 25%. Nanoscribes is an asset for development of next generation genome editing approaches using VLPs. |
| format | Article |
| id | doaj-art-f8b464ff18794d43b99587c3312342b5 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-f8b464ff18794d43b99587c3312342b52025-01-05T12:39:37ZengNature PortfolioNature Communications2041-17232025-01-0116111210.1038/s41467-024-55604-0Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particlesThibaut Halegua0Valérie Risson1Julien Carras2Martin Rouyer3Laurent Coudert4Arnaud Jacquier5Laurent Schaeffer6Théophile Ohlmann7Philippe Emmanuel Mangeot8CIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de LyonPathophysiology and Genetics of Neuron and Muscle, CNRS UMR 5261, INSERM U1315, Université Lyon1, Faculté de Médecine Lyon EstPathophysiology and Genetics of Neuron and Muscle, CNRS UMR 5261, INSERM U1315, Université Lyon1, Faculté de Médecine Lyon EstCIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de LyonPathophysiology and Genetics of Neuron and Muscle, CNRS UMR 5261, INSERM U1315, Université Lyon1, Faculté de Médecine Lyon EstPathophysiology and Genetics of Neuron and Muscle, CNRS UMR 5261, INSERM U1315, Université Lyon1, Faculté de Médecine Lyon EstPathophysiology and Genetics of Neuron and Muscle, CNRS UMR 5261, INSERM U1315, Université Lyon1, Faculté de Médecine Lyon EstCIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de LyonCIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de LyonAbstract Prime Editing can rewrite genes in living cells by allowing point mutations, deletions, or insertion of small DNA sequences with high precision. However, its safe and efficient delivery into human stem cells remains a technical challenge. In this report, we engineer Nanoscribes, virus-like particles that encapsidate ribonucleoprotein complexes of the Prime Editing system and allow their delivery into recipient cells. We identify key features that unlock the potential of Nanoscribes, including the use of multiple fusogens, the improvement of pegRNAs structures, their encoding by a Pol II system and the optimization of Prime-Editors. Nanoscribes edit HEK293T with an efficiency of 68% at the HEK3 locus with increased fidelity over DNA-transfection and support pegRNA-multiplexing. Importantly, Nanoscribes permit editing of myoblasts, hiPSCs and hiPSCs-derived hematopoietic stem cells with an editing efficiency up to 25%. Nanoscribes is an asset for development of next generation genome editing approaches using VLPs.https://doi.org/10.1038/s41467-024-55604-0 |
| spellingShingle | Thibaut Halegua Valérie Risson Julien Carras Martin Rouyer Laurent Coudert Arnaud Jacquier Laurent Schaeffer Théophile Ohlmann Philippe Emmanuel Mangeot Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles Nature Communications |
| title | Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles |
| title_full | Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles |
| title_fullStr | Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles |
| title_full_unstemmed | Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles |
| title_short | Delivery of Prime editing in human stem cells using pseudoviral NanoScribes particles |
| title_sort | delivery of prime editing in human stem cells using pseudoviral nanoscribes particles |
| url | https://doi.org/10.1038/s41467-024-55604-0 |
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