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Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing

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    0524787 - ÚŽFG 2021 RIV GB eng J - Journal Article
    Rohiwal, Sonali Suresh - Dvořáková, N. - Klíma, Jiří - Vaškovičová, Michaela - Šenigl, Filip - Šlouf, Miroslav - Pavlová, Eva - Štěpánek, Petr - Babuka, David - Beneš, Hynek - Ellederová, Zdeňka - Stieger, K.
    Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing.
    Scientific Reports. Roč. 10, č. 1 (2020), č. článku 4619. ISSN 2045-2322. E-ISSN 2045-2322
    R&D Projects: GA MŠMT(CZ) LO1609; GA TA ČR(CZ) TN01000008; GA MŠMT(CZ) LO1507
    Institutional support: RVO:67985904 ; RVO:68378050 ; RVO:61389013
    Keywords : nanoparticles * CRISP/Cas9 * genome editing
    OECD category: Genetics and heredity (medical genetics to be 3); Polymer science (UMCH-V)
    Impact factor: 4.380, year: 2020
    Method of publishing: Open access
    https://www.nature.com/articles/s41598-020-61465-6

    Clustered regularly interspaced short palindromic repeats-associated protein (CRISPR/Cas9) system has become a revolutionary tool for gene editing. Since viral delivery systems have significant side effects, and naked DNA delivery is not an option, the nontoxic, non-viral delivery of CRISPR/Cas9 components would significantly improve future therapeutic delivery. In this study, we aim at characterizing nanoparticles to deliver plasmid DNA encoding for the CRISPR-Cas system in eukaryotic cells in vitro. CRISPR/Cas9 complexed polyethylenimine (PEI) magnetic nanoparticles (MNPs) were generated. We used a stable HEK293 cell line expressing the traffic light reporter (TLR-3) system to evaluate efficient homology- directed repair (HDR) and non-homologous end joining (NHEJ) events following transfection with NPs. MNPs have been synthesized by co-precipitation with the average particle size around 20nmin diameter. The dynamic light scattering and zeta potential measurements showed that NPs exhibited narrow size distribution and sufficient colloidal stability. Genome editing events were as efficient as compared to standard lipofectamine transfection. Our approach tested non-viral delivery of CRISPR/Cas9 and DNA template to perform HDR and NHEJ in the same assay. We demonstrated that PEI-MNPs is a promising delivery system for plasmids encoding CRISPR/Cas9 and template DNA and thus can improve safety and utility of gene editing.
    Permanent Link: http://hdl.handle.net/11104/0309077

     
     
Number of the records: 1  

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