Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Development of Trans-Splicing Aptazyme Which Can Specifically Modify Hepatitis C Virus Genome

C형 간염바이러스(HCV) 유전체를 특이적으로 변형할 수 있는 Trans-Splicing Aptazyme 발굴

  • Kim, Ju-Hyun (Department of Molecular Biology and Institute of Nanosensor and Biotechnology, Dankook University) ;
  • Lee, Chang-Ho (Department of Molecular Biology and Institute of Nanosensor and Biotechnology, Dankook University) ;
  • Jang, Sun-Young (Department of Molecular Biology and Institute of Nanosensor and Biotechnology, Dankook University) ;
  • Lee, Seong-Wook (Department of Molecular Biology and Institute of Nanosensor and Biotechnology, Dankook University)
  • 김주현 (단국대학교 자연과학부 분자생물학과 나노센서 바이오텍 연구소) ;
  • 이창호 (단국대학교 자연과학부 분자생물학과 나노센서 바이오텍 연구소) ;
  • 장선영 (단국대학교 자연과학부 분자생물학과 나노센서 바이오텍 연구소) ;
  • 이성욱 (단국대학교 자연과학부 분자생물학과 나노센서 바이오텍 연구소)
  • Published : 2008.09.30
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Abstract

For the development of specific and effective basic genetic materials to inhibit replication of hepatitis C virus (HCV), HCV genome-targeting trans-splicing aptazyme, which activity is allosterically regulated by a specific ligand, was developed. The aptazyme was designed to be comprised of sequence of RNA aptamer to the ligand, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding the ligand to the aptamer, and trans-splicing ribozyme targeting +199 nt of HCV IRES. Especially, when the aptamer and the communication module was inserted at both P6 and P8 catalytic domain of the specific ribozyme, allosteric activity of the aptazyme was the most induced. The aptazyme was shown to induce activity of trans-splicing reaction specifically and efficiently only in the presence of the specific ligand, but neither in the absence of any ligand nor in the presence of control ligand. This aptazyme can be used as a specific and effective genetic agent against HCV, and a tool for the isolation of anti-HCV lead compounds.

C형 간염바이러스(hepatitis C virus; HCV) 복제를 효과적이며 특이적으로 제어할 수 있는 유전산물을 개발하기 위하여 특정 리간드 존재에 의해 allosteric하게 그 활성이 조절될 수 있는 HCV 유전체 표적 trans-splicing 리보자임(trans-splicing aptazyme)을 발굴하였다. 이러한 trans-splicing aptazyme은 특정 리간드와 특이적으로 결합하는 RNA aptamer 부위, aptamer와 리간드와의 결합에 의해 리보자임 활성을 유도할 수 있도록 구조적 변이를 전달할 수 있는 communication module부위 및 HCV IRES의 +199 nt를 인지하는trans-splicing리보자임 등으로 구성되도록 설계하였다. 특히 trans-splicing 리보자임의 catalytic core의 P6과 P8 부위에 aptamer와 communication module을 삽입하였을 때 가장 allosteric하게 리보자임 활성이 유도되었다. 이러한 리보자임은 리간드가 없거나 대조 리간드가 존재할 때에는 trans-splicing 반응을 유도하지 못하였으나 특정 리간드가 존재할 때에만 효과적이며 특이적으로 trans-splicing 반응을 유도하여 표적 RNA를 변형시킬 수 있음을 관찰하였다. 이러한 aptazyme은 HCV 증식에 대해 특이적이며 효과적인 억제를 위한 선도물질로 이용 가능할 뿐 아니라 HCV 치료선도 물질의 스크리닝용 도구로서도 활용될 수 있을 것이다.

Keywords

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