Article
  • Preparation and Gene Silencing Efficiency of Chitosan-based siRNA Nano-carriers

  • Eunsun Lee*, **, Wheemoon Cho*, Jae Ho Lee***, Sang Jun Park*, Yong-Hee Kim**, and Chun-Ho Kim*,†

  • *Laboratory of Tissue Engineering, Korea Institute of Radiological and Medical Science, 75, Nowon-gil, Nowon-gu, Seoul 01812, Korea
    **Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, 222, Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea
    ***Research Institute for Molecular-Targeted Drugs, Konkuk Institute of Technology, Konkuk University, 120, Neungdong-ro, Gwangjin-gu, Seoul 05029, Korea

  • 키토산 기반 siRNA 나노전달체 제조 및 유전자 억제 효율

  • 이은선*, ** · 조휘문* · 이재호*** · 박상준* · 김용희** · 김천호*,†

  • *한국원자력의학원 방사선의학연구소, **한양대학교 공과대학 생명공학과, ***건국대학교 질병분자표적신약연구소

  • Reproduction, stored in a retrieval system, or transmitted in any form of any part of this publication is permitted only by written permission from the Polymer Society of Korea.

Abstract

This study is to prepare chitosan-based siRNA nano-carriers (Chi-TPP-siRNA) using ionic gelation between cationic chitosan and anionic tripolyphosphate (TPP) for delivery of siRNA to cytoplasm. We studied the effect of the molecular weight of chitosan and the mole ratio of chitosan/TPP on the physicochemical properties of Chi-TPP nanoparticles by using dynamic light scattering and electron microscopy. The siRNA encapsulation was evaluated by using electrophoresis. The biological properties of Chi-TPP-siRNA on MDA-MB-231 cells were evaluated by using confocal laser microscopy. The resulting Chi-TPP-siRNAs, using control siRNA, were formed in sphere shape. The diameter and the PdI value of Chi-TPP-siRNA were 223.5 nm and 0.229, respectively. The cell viability of Chi-TPP-siRNA at the concentration of 500 μg/mL was above 88%. Green fluorescent protein (GFP) silencing effect of Chi-TPP-siGFP was evaluated to be 48.1%. In conclusion, the biocompatible nano-carrier, Chi-TPP-siRNA, could be used in various gene therapy.


세포질로의 siRNA 전달을 위해, 본 연구는 양이온성의 키토산과 음이온성인 tripolyphosphate(TPP)의 이온결합을 이용하여 유전자 나노전달체(Chi-TPP-siRNA)를 제조하였다. 키토산의 분자량 및 키토산과 TPP의 몰비에 따른 나노입자(Chi-TPP)의 물리화학적 특성을 동적광산란기(DLS)와 전자 현미경(EM)으로 확인하였다. siRNA 담지 효과는 전기영동으로 확인하였다. MDA-MB-231에 대한 나노전달체의 생물학적 특성을 공초점현미경으로 확인하였다. 모델 siRNA 담지 Chi-TPP-siRNA 나노전달체는 직경과 다분산도가 각각 223.5 nm, 0.229인 구형으로 형성되었다. 나노전달체 처리군(500 μg/mL)에서의 세포 생존율은 88% 이상이었다. siGFP를 담지한 나노전달체(Chi-TPP-siGFP)는 48.1%의 green fluorescent protein(GFP) 유전자 억제 효율을 보였다. 결과적으로 생체친화성의 Chi-TPP-siRNA 나노전달체는 다양한 유전자 치료에 이용될 수 있다.


Keywords: molecular weight of chitosan, tripolyphosphate, siRNA carrier, monodispersity.

  • Polymer(Korea) 폴리머
  • Frequency : Bimonthly(odd)
    ISSN 0379-153X(Print)
    ISSN 2234-8077(Online)
    Abbr. Polym. Korea
  • 2022 Impact Factor : 0.4
  • Indexed in SCIE

This Article

  • 2021; 45(5): 697-703

    Published online Sep 25, 2021

  • 10.7317/pk.2021.45.5.697
  • Received on Mar 12, 2021
  • Revised on Jun 7, 2021
  • Accepted on Jun 24, 2021

Correspondence to

  • Chun-Ho Kim*

  • *Laboratory of Tissue Engineering, Korea Institute of Radiological and Medical Science, 75, Nowon-gil, Nowon-gu, Seoul 01812, Korea

  • E-mail: chkim@kirams.re.kr