Efficient gene editing via non-viral delivery of CRISPR–Cas9 system using polymeric and hybrid microcarriers

CRISPR–Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarri...

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Bibliographic Details
Published in:Nanomedicine 2018-01, Vol.14 (1), p.97-108
Main Authors: Timin, Alexander S., Muslimov, Albert R., Lepik, Kirill V., Epifanovskaya, Olga S., Shakirova, Alena I., Mock, Ulrike, Riecken, Kristoffer, Okilova, Maria V., Sergeev, Vladislav S., Afanasyev, Boris V., Fehse, Boris, Sukhorukov, Gleb B.
Format: Article
Language:English
Subjects:
CRISPR–Cas9
Gene delivery
Gene editing
Polyelectrolyte microcapsules
Sol–gel
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Summary:CRISPR–Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarriers, made of degradable polymers such as polypeptides and polysaccharides and modified by silica shell, for delivery of all CRISPR–Cas9 components. We found that these microcarriers mediate more efficient transfection than a commercially available liposome-based transfection reagent (>70% vs. 40% vs. 20% for plasmid DNA). For proof-of-concept, we delivered CRISPR–Cas9 components using our capsules to dTomato-expressing HEK293T cells—a model, in which loss of red fluorescence indicates successful gene editing. Notably, transfection of indicator cells translated in high-level dTomato knockout in approx. 70% of transfected cells. In conclusion, we have provided proof-of-principle that our micro-sized containers represent promising non-viral platforms for efficient and safe gene editing. CRISPR–Cas9 is a revolutionary genome-editing technology with huge potential for research, but also clinical applications. Microcapsules combine high loading capacity with easy functionalization, e.g., to allow tissue targeting. Using an indicator cell line we show here that polymeric and hybrid silica-coated capsules mediate efficient, non-toxic and transient delivery of CRISPR–Cas9 system components. Our data suggest that microcapsules represent promising vectors for application of genome-editing tools. [Display omitted]
ISSN:1549-9634
1549-9642
DOI:10.1016/j.nano.2017.09.001