2016 Fiscal Year Research-status Report
Comprehensive identification of cis-regulatory elements involved in the evolutionary origin of the turtle carapace.
| Project/Area Number |
16K18595
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
PASCUAL JUAN 国立研究開発法人理化学研究所, 倉谷形態進化研究室, 研究員 (30594098)
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Keywords | 比較ゲノム / 形態進化 |
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| Outline of Annual Research Achievements |
In order to finely assess the regulatory state of the carapacial ridge, I proposed to perform ATAC sequencing. This technique allows the identification of open chromatin (and thus probably exerting a function) to the nucleotide level. As for the last fiscal year, I was able to collect carapacial ridges as well as limbs and body walls of turtle embryos at stage 14 and was able to successfully sequence the corresponding ATAC libraries. Moreover, I have presented my work related with the origin of the turtle carapace in an international meeting, in China, to which I was invited to give a talk in the World Congress of Herpetology. I also presented a poster in the European Evo-Devo meeting in Sweden.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
My project had two solid phases stated in the proposal: one for the first year, in which I planned to perform ATAC-seq analysis, and another research plan for the second fiscal year, in which I will aim at doing ChIP-seq analysis against turtle Lef1 and beta-catenin. The plan for the first fiscal year has been successfully completed. I was able to construct good ATAC-seq libraries for the tissues mentioned in the previous section, and already got the data from the sequencing provider. At the moment, I am analyzing these data, whose results are promising.
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| Strategy for Future Research Activity |
For the next fiscal year, I will focus on completing the ChIP-seq analysis that I proposed in my Kakenhi project. This will involve seeking for specific antibodies against the turtle Lef1 and beta-catening proteins. In case that commercial antibodies fail, this will mean a delay in the project since I will have to generate custom antibodies, a process that usually takes about 3 months. Fortunately, we are already checking commercial antibodies. Once I have found these antibodies, I will proceed with the ChIP experiment, for which I already have experience from my previous project (histone modifications ChIP-seq analysis).
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| Causes of Carryover |
By means of outsourcing the sequencing, I was able to get the necessary data at a lower price, together with more efficient experiments and less expenditure in business trips.
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| Expenditure Plan for Carryover Budget |
I will be able to sequence even more data in this fiscal year using the remanent money from last one, expectedly up to two lanes of PE 100 data from a HiSeq4000.
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Research Products
(2 results)
