Advancement of revolutional cell technology based on the IR/MAR gene amplification method

2014 Fiscal Year Final Research Report

• Project/Area Number: 23380203
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied molecular and cellular biology
• Research Institution: Hiroshima University
• Principal Investigator: SHIMIZU Noriaki 広島大学, 生物圏科学研究科, 教授 (10216096)
• Project Period (FY): 2011-04-01 – 2015-03-31
• Keywords: 遺伝子増幅 / 細胞工学技術 / 組換え蛋白質生産 / 染色体外遺伝因子 / Double minutes / 細胞癌化

Outline of Final Research Achievements

We previously found that a plasmid bearing mammalian replication initiation region (IR) and a nuclear matrix attachment region (MAR) was amplified spontaneously in animal cells and generate extrachromosomal DMs or chromosomal HSR. This study aimed to develop this method to a novel cell technology. Consequently, 1) we identified minimal required sequence for gene amplification, and clarified the relationship to DNA replication. 2) We clarified the mechanism of gene amplification. 3) We isolated a human genome sequence that enhance the gene expression from tandemly repeated sequence.4) We showed that the pulverization of chromosome de novo generates DMs that had amplified oncogenes. 5) We obtained the understanding about the intracellular maintenance and extracellular elimination of DMs.

Free Research Field

分子細胞生物学