A multiple gene expression switch using a multi-integrase system on a mammalian artificial chromosome vector.

2015 Fiscal Year Final Research Report

• Project/Area Number: 25540134
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Life / Health / Medical informatics
• Research Institution: Tottori University
• Principal Investigator: Ohbayashi Tetsuya 鳥取大学, 生命機能研究支援センター, 准教授 (80348804)
• Co-Investigator(Renkei-kenkyūsha): Nakamura Kazuomi 鳥取大学, 生命機能研究支援センター, プロジェクト研究員 (90598137)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 人工染色体ベクター / インテグレース

Outline of Final Research Achievements

Site-specific recombinases (SSR), such as Cre or Flp recombination target cassettes, have been successfully excised or inverted by a single SSR to regulate transgene expression. This study investigated the potential for expanding the multiple regulation of transgenes using three different integrase systems. We designed three excision cassettes that expressed luciferase, where the luciferase expression could be exchanged to a fluorescent protein by site-specific recombination. Each cassette that was regulated independently by a different integrase was connected in tandem and inserted into a mouse artificial chromosome (MAC) vector in CHO cells. By transient expression of integrase, the targeted luciferase activity was lost and fluorescence was activated. These results suggest that the combined use of these integrase systems in a defined locus on a MAC vector permits the multiple regulation of transgene expression and might contribute to genomic or cell engineering.

Free Research Field

合成生物学