Genome Editing in a Wide Area of the Brain Using Dendrimer-Based Ternary Polyplexes of Cas9 Ribonucleoprotein
| Autor: | Ahmed Fouad Abdelwahab Mohammed, Keiichi Motoyama, Yuki Maeda, Masamichi Inoue, Taishi Higashi, Takuro Niidome, Risako Onodera, Toru Taharabaru, Ryoma Yokoyama |
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| Rok vydání: | 2020 |
| Předmět: |
Dendrimers
Materials science 02 engineering and technology 030226 pharmacology & pharmacy 03 medical and health sciences 0302 clinical medicine Genome editing CRISPR-Associated Protein 9 Cell Line Tumor Dendrimer Animals Humans General Materials Science Ternary complex Ribonucleoprotein Gene Editing Drug Carriers Mice Inbred BALB C Messenger RNA Cas9 Gene Transfer Techniques Brain RNA 021001 nanoscience & nanotechnology Cell biology Ribonucleoproteins Lipofectamine CRISPR-Cas Systems 0210 nano-technology |
| Zdroj: | ACS Applied Materials & Interfaces. 12:21386-21397 |
| ISSN: | 1944-8252 1944-8244 |
| DOI: | 10.1021/acsami.9b21667 |
| Popis: | A preassembled Cas9/single-guide RNA complex (Cas9 ribonucleoprotein; Cas9 RNP) induces genome editing efficiently, with small off-target effects compared with the conventional techniques, such as plasmid DNA and mRNA systems. However, penetration of Cas9 RNP through the cell membrane is low. In particular, the incorporation of Cas9 RNP into neurons and the brain is challenging. In the present study, we have reported the use of a dendrimer (generation 3; G3)/glucuronylglucosyl-β-cyclodextrin conjugate (GUG-β-CDE (G3)) as a carrier of Cas9 RNP and evaluated genome editing activity in the neuron and the brain. A Cas9 RNP ternary complex with GUG-β-CDE (G3) was prepared by only mixing the components. The resulting complex exhibited higher genome editing activity than the complex with the dendrimer (G3), Lipofectamine 3000 or Lipofectamine CRISPRMAX in SH-SY5Y cells, a human neuroblastoma cell line. In addition, GUG-β-CDE (G3) enhanced the genome editing activity of Cas9 RNP in the whole mouse brain after a single intraventricular administration. Thus, GUG-β-CDE (G3) is a useful Cas9 RNP carrier that can induce genome editing in the neuron and brain. |
| Databáze: | OpenAIRE |
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