| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
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| 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.
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