| Budget Amount *help |
¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
|
|---|
| Outline of Final Research Achievements |
I aimed to establish molecular screening for climbing fiber (CF)-Purkinje cell (PC) synapse elimination by knocking out a particular gene with CRISPR/Cas9 system in PCs of mouse whole cerebellum. As a result of testing several conditions, in utero electroporation with a double-electrode probe against both sides of cerebellum at embryonic day 11 showed the highest efficiency of gene transduction into PCs of whole cerebellum. I investigated the effects of knockout of a gene required for CF-PC synapse elimination with CRISPR/Cas9 by electrophysiologically counting the number of CFs on acute cerebellar slices. However, the knockout on PCs by either SpCas9 or eSpCas9 showed no significant impairment on CF-PC synapses. This result suggested a low efficiency of genome editing on PCs in this condition in spite of a high efficiency of gene transduction into whole cerebellar PCs.
|
