Bioluminescence imaging for osteogenic protein secretion

2016 Fiscal Year Final Research Report

• Project/Area Number: 26462828
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Aichi Gakuin University
• Principal Investigator: Suzuki Takahiro 愛知学院大学, 歯学部, 准教授 (70298545)
• Co-Investigator(Renkei-kenkyūsha): INOUYE Satoshi JNC(株), 横浜研究所, 企画研究員 (40426622)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: イメージング / 生物発光 / 開口分泌 / 遺伝子発現 / 3D培養 / 細胞接着 / タンパク質分泌 / ルシフェラーゼ

Outline of Final Research Achievements

A simple method for preparing codon-optimized gene by choosing only codons preferentially used in human genes was named “preferred human codon-optimization method”. We demonstrated that the preferred human codon-optimized luciferase genes were efficiently expressed in several mammalian cells including osteoblastic cells. Also, we expressed a mutated 19 kDa protein of Oplophorus luciferase (nanoKAZ), a preferred codon-optimized gene, and visualized it by bioluminescence imaging. Furthermore, we demonstrated quantitative visualization of protein secretion from 3D-cultured cells by the method of video-rate bioluminescence imaging using Gaussia luciferase as a reporter. Finally, we succeeded in visualizing polarized protein secretion from osteoblastic cells.

Free Research Field

医歯薬学