Spatiotemporal in vivo gene regulation using the electroporation technique

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K15020
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Iwate Medical University
• Principal Investigator: Hitomi Jiro 岩手医科大学, 医学部, 教授 (00218728)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 遺伝子導入 / エレクトロポレーション / ゼブラフィッシュ / Gal4-UAS / 脈管形成 / 血管新生

Outline of Final Research Achievements

We had succeeded in clarifying that the formation of the vascular system of the brain is stepwisely performed in zebrafish during early ontogeny even without the blood flow. To elucidate the regulatory mechanisms, the genes spatiotemporal regulation sytem is necessary, so we challenged to establish the new method for in vivo using a dual pulse electroporation. Firstly, we established a new transgenic zebrafish line Tg(bactin2:GFF), in which Gal4 protein was expressed in the whole embryo. We injected the UAS:EGFP pA plasmid into anywhere in the cranial region of the 2dpf embryo, and then applied the pulse electric potential. As a result, the EGFP expression was observed in the head ubiquitously, that is, we succeeded in controlling the in vivo gene expression spatiotemporally. In future, we will improve the applying the pulse electric potential and achieve higher definitive gene introduction.

Free Research Field

解剖学一般(含組織学・発生学）