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Find and cut-and-transfer (FiCAT) mammalian genome engineering

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dc.contributor.author Pallarès-Masmitjà, Maria
dc.contributor.author Ivančić, Dimitrije
dc.contributor.author Mir-Pedrol, Júlia
dc.contributor.author Jaraba-Wallace, Jessica
dc.contributor.author Tagliani, Tommaso
dc.contributor.author Oliva Miguel, Baldomero
dc.contributor.author Rahmeh, Amal
dc.contributor.author Sánchez-Mejías, Avencia
dc.contributor.author Güell Cargol, Marc
dc.date.accessioned 2022-01-28T08:05:54Z
dc.date.available 2022-01-28T08:05:54Z
dc.date.issued 2021
dc.identifier.citation Pallarès-Masmitjà M, Ivančić D, Mir-Pedrol J, Jaraba-Wallace J, Tagliani T, Oliva B, Rahmeh A, Sánchez-Mejías A, Güell M. Find and cut-and-transfer (FiCAT) mammalian genome engineering. Nat Commun. 2021;12(1):7071. DOI: 10.1038/s41467-021-27183-x
dc.identifier.issn 2041-1723
dc.identifier.uri http://hdl.handle.net/10230/52353
dc.description.abstract While multiple technologies for small allele genome editing exist, robust technologies for targeted integration of large DNA fragments in mammalian genomes are still missing. Here we develop a gene delivery tool (FiCAT) combining the precision of a CRISPR-Cas9 (find module), and the payload transfer efficiency of an engineered piggyBac transposase (cut-and-transfer module). FiCAT combines the functionality of Cas9 DNA scanning and targeting DNA, with piggyBac donor DNA processing and transfer capacity. PiggyBac functional domains are engineered providing increased on-target integration while reducing off-target events. We demonstrate efficient delivery and programmable insertion of small and large payloads in cellulo (human (Hek293T, K-562) and mouse (C2C12)) and in vivo in mouse liver. Finally, we evolve more efficient versions of FiCAT by generating a targeted diversity of 394,000 variants and undergoing 4 rounds of evolution. In this work, we develop a precise and efficient targeted insertion of multi kilobase DNA fragments in mammalian genomes.
dc.description.sponsorship The authors received funding from UPGRADE (European Union Horizon 2020, grant agreement No 825825); Fundació La Caixa: Caixa Impulse 2018 (grant agreement CI 1800036), Caixa Impulse Consolidate 2019 (grant agreement cF91-00010) and Programa Captació de Talent; Fundación Ramón Areces (“Advanced gene editing technologies to restore LAMA2 on merosin-deficient congenital muscular dystrophy type 1A”); Ministerio de Economia, Industria y Competitividad de España (Plan Estatal 2013-2016 (Grant agreement SAF2017-88784-R, Ramón y Cajal program (Grant agreement RYC-2015-17734)). All grants were received by M.G.
dc.format.mimetype application/pdf
dc.language.iso eng
dc.publisher Nature Research
dc.relation.ispartof Nat Commun. 2021;12(1):7071
dc.rights © The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
dc.rights.uri http://creativecommons.org/licenses/by/4.0/
dc.title Find and cut-and-transfer (FiCAT) mammalian genome engineering
dc.type info:eu-repo/semantics/article
dc.identifier.doi http://dx.doi.org/10.1038/s41467-021-27183-x
dc.subject.keyword Genetic engineering
dc.subject.keyword Targeted gene repair
dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/825825
dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/SAF2017-88784-R
dc.rights.accessRights info:eu-repo/semantics/openAccess
dc.type.version info:eu-repo/semantics/publishedVersion

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