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Α Β Γ Δ Ε Ζ Η Θ Ι Κ Λ Μ Ν Ξ Ο Π Ρ Σ Τ Υ Φ Χ Ψ Ω α β γ δ ε ζ η θ ι κ λ μ ν ξ ο π ρ σ τ υ φ χ ψ ω
á à Á À é è É È ç Ç ê
Ä Ö Ü ä ö ü ß
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§ ※ ☆ ★ ○ ● ◎ ◇ ◆ □ ■ △ ▽ → ← ↑ ↓ ↔ 〓 ◁ ◀ ▷ ▶ ♤ ♠ ♡ ♥ ♧ ♣ ⊙ ◈ ▣ ◐ ◑ ▒ ▤ ▥ ▨ ▧ ▦ ▩ ♨ ☏ ☎ ☜ ☞ † ‡ ↕ ↗ ↙ ↖ ↘ ♭ ♩ ♪ ♬ ㉿ ㈜ № ㏇ ™ ㏂ ㏘ ℡ ＃ ＆ ＊ ＠ ª º
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      저자 : Kanjiro Miyata (검색결과 2 건)
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      • Development of Small Polymeric Nanocarriers for Systemic siRNA Delivery

        Kanjiro Miyata,Kazunori Kataoka 한국고분자학회 2014 한국고분자학회 학술대회 연구논문 초록집 Vol.2014 No.10

      • Amphiphilic Polyaspartamide Derivative for mRNA Delivery and their Application for CRISPR-Cas Technology

        Hyun Jin KIM,Satomi OGURA,Kanjiro MIYATA 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10

        Transient and high Cas9 protein expression from messenger RNA (mRNA) is essential for achieving genome engineering or gene editing therapy. To this end, we have developed an amphiphilic polyaspartamide modified with diethylenetriamine (DET) and 2-cyclohexylethyl (CHE) moieties (PAsp(DET/CHE)) [H. J. Kim et al, ACS Cent. Sci. 1866-1875, 2019]. DET moiety provides high endosomal escape ability and CHE moiety optimizes polymer hydrophobicity between mRNA nanoparticle stability in the extracellular condition and mRNA release in the cytoplasm. PAsp(DET/CHE) was synthesized via two-step process. The precursor polymer poly(β-benzyl-L-aspartate) (PBLA) were synthesized with ring opening polymerization between n-butylamine and β-benzyl-L-aspartate N-carboxy-anhydride (BLA-NCA) to have degree of polymerization (DP) of 26. PAsp(DET/CHE) was synthesized by co-aminolysis of DET and CHE into PBLA to contain 15 DET and 11 CHE moieties. PAsp(DET/CHE) produced 150 ㎚-sized nanoparticles with Cas9 mRNA via simple mixing within 30 min. Interestingly, the translation efficacy was not affected by DP of PAsp(DET/CHE) from 26 to 121 but by the amounts of CHE moieties in the single polymer. The PAsp(DET/CHE) with less than 8 CHE introduction ratio did not enhance translation efficacy at all. The PAsp(DET/CHE) polyplex achieved higher S. pyogenes Cas9 (SpCas9) nuclease activity in a luciferase-based homology-directed repair (HDR) assay in cultured cancer cells, compared with the other control polyplexes as well as a commercial transfection reagent control. Ultimately, the PAsp(DET/CHE) nanoparticles locally injected into Ai9 mouse muscle to investigate gene editing efficacy in vivo. The nanoparticles (3 ㎎ Cas9 mRNA and 3 ㎎ sgRNA) successfully induced gene editing effects in the large area of muscle at 3 day post-administration. These results demonstrate the strong potential of PAsp(DET/CHE) for in vitro and in vivo Cas9 mRNA delivery.

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