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

Alexander S. Timin, Albert R. Muslimov, Kirill V. Lepik, Olga S. Epifanovskaya, Alena I. Shakirova, Ulrike Mock, Kristoffer Riecken, Maria V. Okilova, Vladislav S. Sergeev, Boris V. Afanasyev, Boris Fehse, Gleb B. Sukhorukov

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29 Citations (Scopus)

Abstract

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. <50% for mRNA, >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.

Original languageEnglish
Pages (from-to)97-108
Number of pages12
JournalNanomedicine: Nanotechnology, Biology, and Medicine
Volume14
Issue number1
DOIs
Publication statusAccepted/In press - 2017

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Keywords

  • CRISPR-Cas9
  • Gene delivery
  • Gene editing
  • Polyelectrolyte microcapsules
  • Sol-gel

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Molecular Medicine
  • Biomedical Engineering
  • Materials Science(all)
  • Pharmaceutical Science

Cite this

Timin, A. S., Muslimov, A. R., Lepik, K. V., Epifanovskaya, O. S., Shakirova, A. I., Mock, U., Riecken, K., Okilova, M. V., Sergeev, V. S., Afanasyev, B. V., Fehse, B., & Sukhorukov, G. B. (Accepted/In press). Efficient gene editing via non-viral delivery of CRISPR-Cas9 system using polymeric and hybrid microcarriers. Nanomedicine: Nanotechnology, Biology, and Medicine, 14(1), 97-108. https://doi.org/10.1016/j.nano.2017.09.001