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

Pallarès-Masmitjà, Maria; Ivančić, Dimitrije; Mir-Pedrol, Júlia; Jaraba-Wallace, Jessica; Tagliani, Tommaso; Oliva, Baldomero; Rahmeh, Amal; Sánchez-Mejías, Avencia; Güell, Marc


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  <dc:creator>Pallarès-Masmitjà, Maria</dc:creator>
  <dc:creator>Ivančić, Dimitrije</dc:creator>
  <dc:creator>Mir-Pedrol, Júlia</dc:creator>
  <dc:creator>Jaraba-Wallace, Jessica</dc:creator>
  <dc:creator>Tagliani, Tommaso</dc:creator>
  <dc:creator>Oliva, Baldomero</dc:creator>
  <dc:creator>Rahmeh, Amal</dc:creator>
  <dc:creator>Sánchez-Mejías, Avencia</dc:creator>
  <dc:creator>Güell, Marc</dc:creator>
  <dc:date>2021-12-03</dc:date>
  <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>
  <dc:identifier>https://zenodo.org/record/6104372</dc:identifier>
  <dc:identifier>10.1038/s41467-021-27183-x</dc:identifier>
  <dc:identifier>oai:zenodo.org:6104372</dc:identifier>
  <dc:relation>info:eu-repo/grantAgreement/EC/H2020/825825/</dc:relation>
  <dc:relation>url:https://zenodo.org/communities/upgrade-h2020-project</dc:relation>
  <dc:rights>info:eu-repo/semantics/openAccess</dc:rights>
  <dc:rights>https://creativecommons.org/licenses/by/4.0/legalcode</dc:rights>
  <dc:source>Nature Communications 12</dc:source>
  <dc:title>Find and cut-and-transfer (FiCAT) mammalian genome engineering</dc:title>
  <dc:type>info:eu-repo/semantics/article</dc:type>
  <dc:type>publication-article</dc:type>
</oai_dc:dc>
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