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Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing
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SYSNO ASEP 0524787 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing Author(s) Rohiwal, Sonali Suresh (UZFG-Y) ORCID
Dvořáková, N. (CZ)
Klíma, Jiří (UZFG-Y) RID, ORCID
Vaškovičová, Michaela (UZFG-Y) ORCID
Šenigl, Filip (UMG-J) RID
Šlouf, Miroslav (UMCH-V) RID, ORCID
Pavlová, Eva (UMCH-V) RID
Štěpánek, Petr (UMCH-V) RID, ORCID
Babuka, David (UMCH-V)
Beneš, Hynek (UMCH-V) RID, ORCID
Ellederová, Zdeňka (UZFG-Y) RID, ORCID
Stieger, K. (DE)Article number 4619 Source Title Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322
Roč. 10, č. 1 (2020)Number of pages 12 s. Publication form Online - E Language eng - English Country GB - United Kingdom Keywords nanoparticles ; CRISP/Cas9 ; genome editing Subject RIV EB - Genetics ; Molecular Biology OECD category Genetics and heredity (medical genetics to be 3) Subject RIV - cooperation Institute of Macromolecular Chemistry - Macromolecular Chemistry R&D Projects LO1609 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) TN01000008 GA TA ČR - Technology Agency of the Czech Republic (TA ČR) LO1507 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UZFG-Y - RVO:67985904 ; UMG-J - RVO:68378050 ; UMCH-V - RVO:61389013 UT WOS 000520966500036 EID SCOPUS 85081725389 DOI 10.1038/s41598-020-61465-6 Annotation 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. Workplace Institute of Animal Physiology and Genetics Contact Jana Zásmětová, knihovna@iapg.cas.cz, Tel.: 315 639 554 Year of Publishing 2021 Electronic address https://www.nature.com/articles/s41598-020-61465-6
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