Elimination of hepatitis B virus by CRISPR

• Project/Area Number: 26670482
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Infectious disease medicine
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: ITO MASAHIKO 浜松医科大学, 医学部, 助教 (50385423)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: B型肝炎ウイルス / CRISPR/Cas9 / CRISPRi / オカルトHBV / cccDNA / HBV / 潜伏感染 / CRISPR/CAS9 / HBV DNAの切断 / 転写抑制

Outline of Final Research Achievements

Hepatitis B virus (HBV) infection is a major cause of acute and chronic hepatitis and is closely associated with development of cirrhosis and hepatocellular carcinoma (HCC) worldwide. Treatments or drags for the clearance of HBV cccDNA have not been established until now. CRISPR/Cas9 system has possibility to completely eliminate cccDNA in the nuclear of hepatocyte. In this study, we applied CRISPR/Cas9 and CRISPRi system to HBV infected cells, and revealed that these system could repress the transcription of pgRNA and expression of HBs and HBe antigen.

Research Products

• [Journal Article] Involvement of the 3' Untranslated Region in Encapsidation of the Hepatitis C Virus (2016)
  • Author(s): Shi G, Ando T, Suzuki R, Matsuda M, Nakashima K, Ito M, Omatsu T, Oba M, Ochiai H, Kato T, Mizutani T, Sawasaki T, Wakita T, Suzuki T.
  • Journal Title: PLoS Pathog Volume: 12(2) Issue: 2 Pages: e1005441-e1005441
  • DOI: 10.1371/journal.ppat.1005441
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] CRISPRシステムによるB型肝炎ウイルスゲノムの排除と転写抑制 (2015)
  • Author(s): 伊藤昌彦, 鈴木哲朗.
  • Journal Title: 月刊細胞 Volume: 47 Pages: 566-570
  • NAID: 40020648478
  • Peer Reviewed
• [Journal Article] Involvement of Hepatitis C Virus NS5A Hyperphosphorylation Mediated by Casein Kinase I-α in Infectious Virus Production. (2014)
  • Author(s): Masaki T, Matsunaga S, Takahashi H, Nakashima K, Kimura Y, Ito M, Matsuda M, Murayama A, Kato T, Hirano H, Endo Y, Lemon SM, Wakita T, Sawasaki T, Suzuki T
  • Journal Title: J Virol Volume: 88(13) Issue: 13 Pages: 7541-55
  • DOI: 10.1128/jvi.03170-13
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] HCV infection upregulates expression of cytosolic phospholipase A2 gamma via activation of c-Myc and NFkB pathways. (2015)
  • Author(s): Ito M，Fukasawa M，Kohara M，Suzuki T.
  • Organizer: 第63回日本ウイルス学会学術集会
  • Place of Presentation: 福岡市
  • Year and Date: 2015-11-22
• [Presentation] Lipidomic analysis of the livers in HCV-infected humanized mice by imaging mass spectrometry. (2015)
  • Author(s): Ito M，Fukasawa M，Kohara M，Suzuki T.
  • Organizer: 第22回C型肝炎及び関連ウイルスに関する国際会議(HCV2015)
  • Place of Presentation: フランス・ストラスブール
  • Year and Date: 2015-10-10
  • Int'l Joint Research
• [Presentation] LUC7L3/Luc7A/CROP as a negative regulator of enhancer II/basal core promoter of hepatitis B virus. (2015)
  • Author(s): Ito M, Li Y, Sun S, Nakashima K, Suzuki T.
  • Organizer: 2015 International Meeting on Molecular Biology of Hepatitis B Viruses.
  • Place of Presentation: ドイツ・バードナウハイム
  • Year and Date: 2015-10-06
  • Int'l Joint Research
• [Presentation] HuH-7由来オーバル細胞様細胞株HdoによるHCV 生活環の調節に関わる分化関連宿主因子の同定. (2014)
  • Author(s): 伊藤昌彦、福原崇介、鈴木亮介、田川陽一、松浦善治、脇田隆字、鈴木哲朗.
  • Organizer: 第62回日本ウイルス学会学術集会.
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-10 – 2014-11-12
• [Presentation] Use of HuH7-derived, bidirectional oval-like cells to identify differentiation dependent host factors that are involved in regulation of HCV lifecycle. (2014)
  • Author(s): Ito M, Fukuhara T, Suzuki R, Matsuura Y, Wakita T, Suzuki T.
  • Organizer: 21st International Symposium on Hepatitis C and Related Viruses.
  • Place of Presentation: カナダ・バンフ
  • Year and Date: 2014-09-07 – 2014-09-11
• [Presentation] Identification of novel factors that are implicated in transcriptional regulation of Hepatitis B virus pregenome. (2014)
  • Author(s): Ito M, Li Y, Sun S, Suzuki T.
  • Organizer: 2014 International Meeting on Molecular Biology of Hepatitis B Viruses.
  • Place of Presentation: アメリカ合衆国・ロサンゼルス
  • Year and Date: 2014-09-03 – 2014-09-06