Elucidation of virus multiplication mechanism by a novel hepatitis B virus replication system using stabilized polymerase

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K07515
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49060:Virology-related
• Research Institution: Hiroshima University
• Principal Investigator: Sakaguchi Takemasa 広島大学, 医系科学研究科(医), 教授 (70196070)
• Co-Investigator(Kenkyū-buntansha): 東浦 彰史 広島大学, 医系科学研究科(医), 助教 (90598129)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: B型肝炎ウイルス / ポリメラーゼ / センダイウイルス / プレゲノムRNA

Outline of Final Research Achievements

Hepatitis B is a serious public health issue. We discovered stabilization occurs when a tag is added to the Hepatitis B polymerase. Taking advantage of this, a system was developed to amplify genomic DNA using a Sendai virus expressing HBV pre-genomic RNA and a stable polymerase expression plasmid. However, complete replication of HBV was not observed. Additionally, we could not achieve mass production or protein crystallization of the HBV polymerase using either the Sendai virus vector or Expi293 cells. The stabilization of the HBV polymerase was insufficient, and thus the outcomes of our research were limited.

Free Research Field

ウイルス学

Academic Significance and Societal Importance of the Research Achievements

SeV-pgRNAとHBVポリメラーゼを用いた系で、SeVの広い宿主域からHBVの肝由来細胞特異的増殖のための因子が明らかになると考えられる。また、これらを改変してSeVのみでHBV増殖が起こせるようになれば、免疫が正常なマウスに投与して肝炎の研究を行うことができる可能性がある。さらにHBVのポリメラーゼの立体構造の解明によって酵素としての作用メカニズムが正確に明らかになれば、ポリメラーゼを標的とした薬剤の開発に資する。