Establishment of a new method using pluripotent stem cells and RNA interference and examination of the Htt gene by the method

2015 Fiscal Year Final Research Report

• Project/Area Number: 25670428
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurology
• Research Institution: National Center of Neurology and Psychiatry
• Principal Investigator: Hohjoh Hirohiko 国立研究開発法人国立精神・神経医療研究センター, 神経研究所神経薬理研究部, 室長 (60238722)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 神経変性疾患 / ハンチントン病 / ハンチンチン遺伝子 / RNAi / shRNA / AAV

Outline of Final Research Achievements

The role and function of huntingtin (Htt) in cells has been poorly understood. In this study we attempted to obtain novel findings to lead to the elucidation of the Htt functions by a new approach using pluripotent stem cells and a RNA interference (RNAi) technique. Optimal small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) against Htt have been successfully designed and shRNA-expression adeno-associated viruses (AAVs) have been constructed. Transmission of the constructed shRNA-expression AAV to mouse ES cells for suppressing the endogenous Htt genes was carried out, but resulted in a failure with a pretty low efficiency of AAV infection. A conventional method with synthetic siRNAs and a transfection reagent was also performed; however, the results could not indicate any marked suppression of the target Htt. Accordingly, the research project was unexpectedly difficult; but, it might be achieved if a recent genome-editing technique such as a CRISPR/Cas9 system were used.

Free Research Field

分子生命科学