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

Research output: Contribution to journalArticle

17 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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Clustered Regularly Interspaced Short Palindromic Repeats
Transfection
Genes
Inborn Genetic Diseases
Liposomes
Polypeptides
Polysaccharides
Silicon Dioxide
Capsules
Containers
Polymers
Plasmids
DNA
Fluorescence
Silica
Technology
Peptides
Gene Editing
Therapeutics

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

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

In: Nanomedicine: Nanotechnology, Biology, and Medicine, Vol. 14, No. 1, 2017, p. 97-108.

Research output: Contribution to journalArticle

Timin, AS, Muslimov, AR, Lepik, KV, Epifanovskaya, OS, Shakirova, AI, Mock, U, Riecken, K, Okilova, MV, Sergeev, VS, Afanasyev, BV, Fehse, B & Sukhorukov, GB 2017, 'Efficient gene editing via non-viral delivery of CRISPR-Cas9 system using polymeric and hybrid microcarriers', Nanomedicine: Nanotechnology, Biology, and Medicine, vol. 14, no. 1, pp. 97-108. https://doi.org/10.1016/j.nano.2017.09.001
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. / Efficient gene editing via non-viral delivery of CRISPR-Cas9 system using polymeric and hybrid microcarriers. In: Nanomedicine: Nanotechnology, Biology, and Medicine. 2017 ; Vol. 14, No. 1. pp. 97-108.
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