Genome Editing in Primary Cells and in Vivo Using Viral-Derived Nanoblades Loaded with Cas9-SgRNA Ribonucleoproteins
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Abstract
Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. However, their delivery into target cells can be technically challenging when working with primary cells or in vivo. Here, we use engineered murine leukemia virus-like particles loaded with Cas9-sgRNA ribonucleoproteins (Nanoblades) to induce efficient genome-editing in cell lines and primary cells including human induced pluripotent stem cells, human hematopoietic stem cells and mouse bone-marrow cells. Transgene-free Nanoblades are also capable of in vivo genome-editing in mouse embryos and in the liver of injected mice. Nanoblades can be complexed with donor DNA for “all-in-one” homology-directed repair or programmed with modified Cas9 variants to mediate transcriptional up-regulation of target genes. Nanoblades preparation process is simple, relatively inexpensive and can be easily implemented in any laboratory equipped for cellular biology.
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Murine-leukemia-based VLPs carrying Cas9-gRNA RNP (nanoblades) showed 67% editing efficiency in human-induced pluripotent stem cells targeting the EMX1 gene.
“Another study developed murine-leukemia-based VLPs carrying the Cas9-gRNA RNP complex (named nanoblades) and showed 67% editing efficiency in human-induced pluripotent stem cells (hiPSCs) targeting the EMX1 gene.”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [84]
In mouse liver, nanoblades showed 7-13% editing efficiency.
“In the mouse liver, they were able to show 7-13% editing efficiency [84].”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [84]
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- Authors
- Mangeot, P. E., Risson, V., Fusil, F., Marnef, A., Laurent, E., Blin, J., Mournetas, V., Massouridès, E., Sohier, T. J. M., Corbin, A., Aubé, F., Teixeira, M., Pinset, C., Schaeffer, L., Legube, G., Cosset, F.-L., Verhoeyen, E., Ohlmann, T., & Ricci, E. P.
- Affiliations
- Lyon 1 Université (FR)
- Centre National de la Recherche Scientifique (FR)
- Inserm (FR)
- Corresponding author
- Mangeot, Philippe E.
- Ricci, Emiliano P.
- ORCID
- Venue
- Nature Communications
- Published
- 2019-01-03
- Volume
- 10
- Issue
- 1
- Pages
- 45-45
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC6318322
- OpenAlex
- W2949383259
- Cited by
- 346
- References
- 56
- Citations by year
- 2026: 37 · 2025: 55 · 2024: 60 · 2023: 48 · 2022: 61 · 2021: 42 · 2020: 26 · 2019: 16 · 2012: 1
- FWCI
- 7.47
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- Europe PMC (PubMed Central) (publishedVersion, other-oa)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- HAL (submittedVersion, other-oa)
- HAL (submittedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- Styrelsen för Internationellt Utvecklingssamarbete
- Fondation Innovations en Infectiologie
- European Commission (HORIZON2020)
- Agence Nationale de la Recherche (ANR-10-INBS-0009, ANR-11-IDEX-0007, ANR-11-LABX-0048, ANR-11-IDEX, ANR-10, IDEX-0007, 11 IDEX - 0007)
- Institut National de la Santé et de la Recherche Médicale (ANR-11-IDEX-0007)
- Centre National de la Recherche Scientifique
- Université Claude Bernard Lyon 1 (ANR-11-IDEX-0007, ANR-11-LABX-0048)
- Université de Lyon (ANR-11-IDEX-0 0 07, ANR-11-LABX-0048)
- École Normale Supérieure de Lyon
- Laboratoire d'Excellence Ecofect (ANR-11-IDEX-0007, ANR-11-LABX-0048)
- H2020 European Research Council (805500)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- genome editing, primary cells, in vivo gene editing, induced pluripotent stem cells, hematopoietic stem cells, bone marrow, homology-directed repair, transcriptional upregulation, mouse embryo
- MeSH terms
- Gene Editing, CRISPR-Associated Protein 9, Animals, DNA Repair, Embryo, Mammalian, Fibroblasts, Genetic Vectors, Hematopoietic Stem Cells, Humans, Macrophages, Mice, Inbred C57BL, Leukemia Virus, Murine, Ribonucleoproteins, Transcriptional Activation, Genome, Cell Line, Tumor, Mice, Induced Pluripotent Stem Cells, HEK293 Cells, Primary Cell Culture
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