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, 1982-
  • dc.date.accessioned 2022-01-28T08:05:54Z
  • dc.date.available 2022-01-28T08:05:54Z
  • dc.date.issued 2021
  • 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.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.doi http://dx.doi.org/10.1038/s41467-021-27183-x
  • dc.identifier.issn 2041-1723
  • dc.identifier.uri http://hdl.handle.net/10230/52353
  • dc.language.iso eng
  • dc.publisher Nature Research
  • dc.relation.ispartof Nat Commun. 2021;12(1):7071
  • dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/825825
  • dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/SAF2017-88784-R
  • 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.accessRights info:eu-repo/semantics/openAccess
  • dc.rights.uri http://creativecommons.org/licenses/by/4.0/
  • dc.subject.keyword Genetic engineering
  • dc.subject.keyword Targeted gene repair
  • dc.title Find and cut-and-transfer (FiCAT) mammalian genome engineering
  • dc.type info:eu-repo/semantics/article
  • dc.type.version info:eu-repo/semantics/publishedVersion

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