Journal article Open Access
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
DCAT Export
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<dct:title>Find and cut-and-transfer (FiCAT) mammalian genome engineering</dct:title>
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<dct:description><p><strong>Abstract</strong></p> <p>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.</p></dct:description>
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Indexed in
- Publication date:
- December 3, 2021
- DOI:
-
- Published in:
- Nature Communications: 12.
- Grants:
-
European Commission:
- UPGRADE - Unlocking Precision Gene Therapy (825825)
- Communities:
- License (for files):
- Creative Commons Attribution 4.0 International