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dc.contributor.authorLostalé Seijo, Irene
dc.contributor.authorLouzao Pernas, Iria
dc.contributor.authorJuanes Carrasco, María Luisa
dc.contributor.authorMontenegro García, Javier
dc.date.accessioned2020-05-26T18:44:48Z
dc.date.available2020-05-26T18:44:48Z
dc.date.issued2017
dc.identifier.citationLostalé-Seijo, I., Louzao, I., Juanes, M., & Montenegro, J. (2017). Peptide/Cas9 nanostructures for ribonucleoprotein cell membrane transport and gene edition. Chemical Science, 8(12), 7923-7931. https://doi.org/10.1039/c7sc03918b
dc.identifier.issn2041-6520
dc.identifier.urihttp://hdl.handle.net/10347/22611
dc.description.abstractThe discovery of RNA guided endonucleases has emerged as one of the most important tools for gene edition and biotechnology. The selectivity and simplicity of the CRISPR/Cas9 strategy allows the straightforward targeting and editing of particular loci in the cell genome without the requirement of protein engineering. However, the transfection of plasmids encoding the Cas9 and the guide RNA could lead to undesired permanent recombination and immunogenic responses. Therefore, the direct delivery of transient Cas9 ribonucleoprotein constitutes an advantageous strategy for gene edition and other potential therapeutic applications of the CRISPR/Cas9 system. The covalent fusion of Cas9 with penetrating peptides requires multiple incubation steps with the target cells to achieve efficient levels of gene edition. These and other recent reports suggested that covalent conjugation of the anionic Cas9 ribonucleoprotein to cationic peptides would be associated with a hindered nuclease activity due to undesired electrostatic interactions. We here report a supramolecular strategy for the direct delivery of Cas9 by an amphiphilic penetrating peptide that was prepared by a hydrazone bond formation between a cationic peptide scaffold and a hydrophobic aldehyde tail. The peptide/protein non-covalent nanoparticles performed with similar efficiency and less toxicity than one of the best methods described to date. To the best of our knowledge this report constitutes the first supramolecular strategy for the direct delivery of Cas9 using a penetrating peptide vehicle. The results reported here confirmed that peptide amphiphilic vectors can deliver Cas9 in a single incubation step, with good efficiency and low toxicity. This work will encourage the search and development of conceptually new synthetic systems for transitory endonucleases direct delivery.
dc.description.sponsorshipThis work was partially supported by the Spanish Agencia Estatal de Investigación (AEI) [CTQ2014-59646-R], the Xunta de Galicia (ED431G/09 and 2016-AD031) and the ERDF. M. J. received a F. P. I. fellowship from MINECO. J. M. received a Ramón y Cajal (RYC-2013-13784), an ERC Starting Investigator Grant (DYNAP-677786) and a Young Investigator Grant from the Human Frontier Science Research Program (RGY0066/2017)
dc.language.isoeng
dc.publisherRoyal Society of Chemistry
dc.relationinfo:eu-repo/grantAgreement/EC/H2020/677786
dc.relationinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2014-59646-R/ES/NUEVOS DISEÑOS CON CONTROL TOPOLOGICO DE PEPTIDOS PENETRANTES EN CELULAS
dc.relationinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/RYC-2013-13784/ES
dc.rights© The Royal Society of Chemistry 2017. This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence.
dc.rights.urihttps://creativecommons.org/licenses/by-nc/3.0/
dc.titlePeptide/Cas9 nanostructures for ribonucleoprotein cell membrane transport and gene edition
dc.typeinfo:eu-repo/semantics/article
dc.identifier.DOI10.1039/c7sc03918b
dc.relation.publisherversionhttps://doi.org/10.1039/C7SC03918B
dc.type.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.e-issn2041-6539
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.contributor.affiliationUniversidade de Santiago de Compostela. Centro de Investigación en Química Biolóxica e Materiais Moleculares
dc.contributor.affiliationUniversidade de Santiago de Compostela. Departamento de Química Orgánica
dc.description.peerreviewedSI


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© The Royal Society of Chemistry 2017. This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence.
Except where otherwise noted, this item's license is described as  © The Royal Society of Chemistry 2017. This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence.





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