Establishment of a synthetic promoter-based system for sensing oncogenic alteration in human live thyroid cells at a single-cell level

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K09438
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Endocrinology
• Research Institution: Nagasaki University
• Principal Investigator: SAENKO Vladimir 長崎大学, 原爆後障害医療研究所, 准教授 (30343346)
• Co-Investigator(Kenkyū-buntansha): ログノビッチ タチアナ 長崎大学, 原爆後障害医療研究所, 助教 (30423643) ; 光武 範吏 長崎大学, 原爆後障害医療研究所, 准教授 (50404215)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 甲状腺癌 / 癌遺伝子 / 遺伝子プロモーター / 合成生物学 / 単一細胞レポーター

Outline of Final Research Achievements

Establishment of a system for sensing oncogene activation at a single-cell level in live cells was the purpose. The system employs a fluorescent reporter driven by a synthetic promoter consisting of transcription factor binding sites (TFBSs) that correlated with upregulation of gene expression after oncogenic challenge. The library of single- or multiple-copy TFBSs was prepared and screened for activity in a dual-luminescent assay. The best promoters were inserted into the fluorescent reporter plasmid and screened by cotransfection with small amounts of oncogene-encoding vectors. Several selected synthetic promoter-reporter plasmids were stably transfected into target cells, which after transfection with oncogene-encoding vectors displayed a fluorescent signal observed by live cell imaging.

Free Research Field

分子生物学