Establishment of Purkinje cell specific CRISPR/Cas9 screening in whole cerebellum
| Project/Area Number |
16K14553
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Neurophysiology / General neuroscience
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
Watanabe Takaki 東京大学, 大学院医学系研究科(医学部), 特任助教 (90749798)
|
|---|
| Project Period (FY) |
2016-04-01 – 2018-03-31
|
| Project Status |
Completed (Fiscal Year 2017)
|
| 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)
|
|---|
| Keywords | 小脳 / プルキンエ細胞 / 登上線維 / CRISPR/Cas9 / シナプス / シナプス刈り込み / 神経科学 |
|---|
| 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.
|
Report
(3 results)
Research Products
(8 results)
-
-
-
[Journal Article] Alternative splicing in the C-terminal tail of Cav2.1 is essential for preventing a neurological disease in mice.2017
Author(s)
Aikawa T, Watanabe T, Miyazaki T, Mikuni T, Wakamori M, Sakurai M, Aizawa H, Ishizu N, Watanabe M, Kano M, Mizusawa H, Watase K.
-
Journal Title
Hum Mol Genet.
Volume: 26(16)
Pages: 3094-3104
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
-
-
-
-
-
