Mostrar el registro sencillo del ítem
dc.contributor.author
Gutierrez, Alejandra
dc.contributor.author
Abrey Recalde, Maria Jimena
dc.contributor.author
Mangeot, Philippe E.
dc.contributor.author
Costa, Caroline
dc.contributor.author
Bernandin, Ornellie
dc.contributor.author
Fusil, Floriane
dc.contributor.author
Froment, Gisèle
dc.contributor.author
Martin, Francisco
dc.contributor.author
Bellabdelah, Karim
dc.contributor.author
Ricci, Emiliano P.
dc.contributor.author
Ayuso, Eduard
dc.contributor.author
Cosset, François loic
dc.contributor.author
Verhoeyen, Els
dc.date.available
2021-07-12T13:39:02Z
dc.date.issued
2019
dc.identifier.citation
Site Specific Knock-In Genome Editing in Human HSCs Using Baboon Envelope gp Pseudotypedviral Derived “Nanoblades” Loaded with Cas9/sgRNA Combined with Donor Encoding AAV-6; American Society of Cell and Gene Therapy 22nd Annual Metting; Washington; Estados Unidos; 2019
dc.identifier.issn
1525-0016
dc.identifier.uri
http://hdl.handle.net/11336/135844
dc.description.abstract
Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. Here, we have designed ?Nanoblades?, a new technology that will deliver a genomic cleaving agent into cells. These are genetically modified Murine Leukemia Virus (MLV) or HIV derived virus like particle (VLP), in which the viral structural protein Gag has been fused to the Cas9. These VLPs are thus loaded with Cas9 protein together with the guide RNAs. Thus, nanoblades are devoid of any viral-derived genetic material. Highly efficient gene editing was obtained in cell lines, IPS cells and primary mouse and human cells (Mangeot et al. Nature Communication, 2019). However, their delivery into target cells can be technically challenging when working with primary immune cells. Now we showed that nanoblades were remarkably efficient for entry into human T, B and hematopoietic stem cells thanks to their surface co-pseudotyping with baboon retroviral and VSVG envelope glycoproteins. We were able to induce efficient, transient and very rapidlygenome-editing in human induced pluripotent stem cells reaching up to 70% in the empty spiracles homeobox 1 (EMX1) and muscular dystrophy (MD) gene locus. A brief nanoblade incubation of primary human T and B cells resulted in 40% and 20% editing of the Wiskott-Aldrich syndrome (WAS) gene locus, while hematopoietic stem cells treated for 18 h with nanoblades allowed 30-40% gene editing in the WAS gene locus and up to 80% for the Myd88 genomic target. Moreover, for the HIV- and MLV-derived nanoblades no cell toxicity and low to undetectable off-target effects were demonstrated.Finally, we also treated hHSCs with nanoblades in combination with an AAV-6 donor encoding vector resulting in over 20% of stable expression cassette knock-in into the WAS gene locus. Currently, we are evaluating these gene modified HSCs for their long-term reconstitution of NOD/SCIDgC-/- mice.Summarizing, this new technology is simple to implement in any laboratory, shows high flexibility for different targets including primary immune cells of murine and human origin, is relatively inexpensive and therefore have important prospects for basic and clinical translation in the area of gene therapy.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
NANOBLADES
dc.subject
GENE EDITING
dc.subject
CRISPR
dc.subject.classification
Biotecnología relacionada con la Salud
dc.subject.classification
Biotecnología de la Salud
dc.subject.classification
CIENCIAS MÉDICAS Y DE LA SALUD
dc.title
Site Specific Knock-In Genome Editing in Human HSCs Using Baboon Envelope gp Pseudotypedviral Derived “Nanoblades” Loaded with Cas9/sgRNA Combined with Donor Encoding AAV-6
dc.type
info:eu-repo/semantics/publishedVersion
dc.type
info:eu-repo/semantics/conferenceObject
dc.type
info:ar-repo/semantics/documento de conferencia
dc.date.updated
2021-07-07T15:17:21Z
dc.journal.pais
Estados Unidos
dc.journal.ciudad
Buenos Aires
dc.description.fil
Fil: Gutierrez, Alejandra. Inserm; Francia
dc.description.fil
Fil: Abrey Recalde, Maria Jimena. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; Argentina. Inserm; Francia
dc.description.fil
Fil: Mangeot, Philippe E.. Inserm; Francia
dc.description.fil
Fil: Costa, Caroline. Inserm; Francia
dc.description.fil
Fil: Bernandin, Ornellie. Inserm; Francia
dc.description.fil
Fil: Fusil, Floriane. Inserm; Francia
dc.description.fil
Fil: Froment, Gisèle. Inserm; Francia
dc.description.fil
Fil: Martin, Francisco. Inserm; Francia
dc.description.fil
Fil: Bellabdelah, Karim. Universidad de Granada; España
dc.description.fil
Fil: Ricci, Emiliano P.. Inserm; Francia
dc.description.fil
Fil: Ayuso, Eduard. Universite de Nantes; Francia
dc.description.fil
Fil: Cosset, François loic. Inserm; Francia
dc.description.fil
Fil: Verhoeyen, Els. Inserm; Francia
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://asgct.org/global/documents/asgct19_abstracts_-final
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.coverage
Internacional
dc.type.subtype
Reunión
dc.description.nombreEvento
American Society of Cell and Gene Therapy 22nd Annual Metting
dc.date.evento
2019-04
dc.description.ciudadEvento
Washington
dc.description.paisEvento
Estados Unidos
dc.type.publicacion
Journal
dc.description.institucionOrganizadora
American Society of Cell and Gene Therapy
dc.source.revista
Molecular Therapy
dc.type
Reunión
Archivos asociados