4.8 Article

Graphene oxide-mediated Cas9/sgRNA delivery for efficient genome editing

Journal

NANOSCALE
Volume 10, Issue 3, Pages 1063-1071

Publisher

ROYAL SOC CHEMISTRY
DOI: 10.1039/c7nr07999k

Keywords

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Funding

  1. National Natural Science Foundation of China [21475048]
  2. National Science Fund for Distinguished Young Scholars of Guangdong Province [2014A030306008]
  3. Project of Guangzhou Science and Technology Plan [201508020003]
  4. Special Support Program of Guangdong Province [2014TQ01R599]
  5. Outstanding Young Teacher Training Program of Guangdong Province [HS2015004]

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Direct cellular delivery of CRISPR/Cas9 complexes is of great significance for genome editing and other recently developed applications, such as gene expression regulation and RNA/DNA imaging. Here, we first constructed a graphene oxide (GO)-polyethylene glycol (PEG)-polyethylenimine (PEI) nanocarrier for the delivery of high-molecular-weight Cas9/single-guide RNA (sgRNA) complexes for endocytosis, endosomal escape, nuclear entry, and gene editing. The results demonstrate that the nanocarrier can be used successfully for efficient gene editing in human AGS cells with an efficiency of similar to 39%. The results also show that this nanocarrier can protect sgRNA from enzymatic degradation, thus exhibiting extremely high stability, which is critical for future in vivo applications. Thus, this GO-mediated Cas9/sgRNA delivery system has potential as a new approach for biomedical research and targeted gene engineering applications.

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