Article
  • Synthesis and Characterization of KKRK Peptide-PAMAM Dendrimer G2 Conjugates as Gene Carriers
  • Younjin Kim and Joon Sig Choi

  • Department of Biochemistry, College of Natural Sciences, Chungnam National University, 99 Daehak-ro, Daejeon 34134, Korea

  • 2세대의 PAMAM 덴드리머와 KKRK 펩티드를 접합한 고분자 소재의 합성 및 유전자 전달체 특성 연구
  • 김연진 ·최준식

  • 충남대학교 자연과학대학 생화학과

  • 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

In this study, new hybrid polymers composed of polyamidoamine dendrimer generation 2 (PAMAM G2) and KKRK, KKRKH peptides derived from a nuclear localization sequence (NLS) were synthesized and characterized as gene carriers. Using Hep G2 and NIH3T3 cell lines, it was confirmed that the gene transfection efficiency was increased about 20- and 2700-fold in each cell compared to that of the native PAMAM G2. The main endocytosis mechanism of the polymer/plasmid DNA complex (polyplex) was proved to be macropinocytosis through endocytosis inhibition experiments using various inhibitors. It was confirmed that the polyplexes existed around the cell nucleus using a confocal microscope. In addition, it was observed that the polyplex showed active escape from endosomes using FACS analysis using acridine orange. As a result, KKRK-PAMAM G2 and KKRKH-PAMAM G2 show their potential to be one of efficient candidates as non-viral vectors.


본 연구에서, 2세대의 폴리도아민 덴드리머(PAMAM G2)와 nuclear localization sequence(NLS) 서열을 최적화한 KKRK, KKRKH 펩티드를 이용하여 새로운 하이브리드 고분자를 합성하였고 이들의 유전자 전달체로서의 특성을 확인하였다. Hep G2와 NIH3T3 세포주를 이용하여 유전자 형질변환 실험으로, 원래의 PAMAM G2 고분자와 비교하여 각 세포에서 20배, 2700배 이상 증가된 유전자 전달 효율을 확인하였다. 다양한 세포 내 이입 저해제들을 이용한 실험을 통해 본 고분자/플라스미드 DNA 복합체의 주된 엔도사이토시스 경로가 거대음작용 메카니즘임을 확인하였다. 공초점 현미경을 통해 플라스미드 DNA가 세포 핵 주변에 분포함을 확인하였다. 또한, FACS 실험으로 고분자/DNA 복합체인 폴리플렉스의 엔도좀의 탈출 효과가 활발함을 확인하였다. 본 연구를 통해 KKRK-PAMAM G2, KKRKH-PAMAM G2가 효율적인 비바이러스성 벡터의 후보군으로 활용될 수 있는 잠재적 가능성을 확인하였다.


Keywords: polyamidoamine dendrimer, nuclear localization sequence, cytotoxicity, endocytosis, transfection.

  • Polymer(Korea) 폴리머
  • Frequency : Bimonthly(odd)
    ISSN 0379-153X(Print)
    ISSN 2234-8077(Online)
    Abbr. Polym. Korea
  • 2023 Impact Factor : 0.4
  • Indexed in SCIE

This Article

  • 2022; 46(1): 113-121

    Published online Jan 25, 2022

  • 10.7317/pk.2022.46.1.113
  • Received on Oct 14, 2021
  • Revised on Oct 29, 2021
  • Accepted on Nov 4, 2021

Correspondence to

  • Joon Sig Choi
  • Department of Biochemistry, College of Natural Sciences, Chungnam National University, 99 Daehak-ro, Daejeon 34134, Korea

  • E-mail: joonsig@cnu.ac.kr