Novel animal models for hereditary diseases using human hepatocytes transplanted chimeric mice and vaculovirus

2016 Fiscal Year Final Research Report

• Project/Area Number: 26420796
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Hiroshima University
• Principal Investigator: Chayama Hiromi 広島大学, 医歯薬保健学研究院(医), 准教授 (70572329)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ゲノム編集 / ヒト肝細胞キメラマウス / 遺伝性肝疾患

Outline of Final Research Achievements

In this study, I established two type of CRISPR-Cas9 systems using vaculovirus vectors that work to knock out or mutate causative genes of hemochromatosis, porphyria, glycogenosis, amino acid metabolism abnormality, urea cycle disorder, constitutional jaundice, fibrotic cystic liver disease and Wilson’s disease. I transfected theses constructs into HEK293 cells and harvested vaculoviruses. However, when I inoculated HuH7 cells and HepG2 cells with the vaculoviruses, the cell growths’ were seriously inhibited. From these results, I decided to improve vaculovirus vectors. Now I confirming whether the targeted genome editing was occurred using the improved CRISPR-Cas9 systems. Further experiments are necessary. I will establish in vitro hereditary liver diseases models using human primary hepatocytes derived from human hepatocytes transplanted chimeric mice and in vivo models using human hepatocytes transplanted chimeric mice with these CRISPR-Cas9 systems.

Free Research Field

消化器病