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Simple and Fast Rolling Circle Amplification-Based Detection of Topoisomerase 1 Activity in Crude Biological Samples

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Gene amplification system based on double rolling-circle replication as a model for oncogene-type amplification.

Takaaki Watanabe¹, Hideyuki Tanabe, Takashi Horiuchi

¹National Institute for Basic Biology, Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Myodaiji, Okazaki, Aichi, 444-8585, Japan. watatka@nibb.ac.jp

Nucleic Acids Research

|June 10, 2011

Summary

Researchers developed a novel gene amplification system using double rolling-circle replication (DRCR) and the Cre-lox system. This model mimics oncogene and drug-resistance gene amplification seen in cancer, aiding further research.

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Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA

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Published on: June 25, 2014

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Gene amplification is crucial in cancer progression and drug resistance.
The precise molecular mechanisms underlying gene amplification are not fully understood.

Purpose of the Study:

To develop a novel system for studying gene amplification in yeast and mammalian cells.
To investigate the molecular mechanisms of gene amplification, particularly in the context of oncogenes and drug resistance.

Main Methods:

Development of a gene amplification system based on double rolling-circle replication (DRCR).
Utilized the Cre-lox system to induce DRCR through a combination of recombination and replication.
Applied the system in both yeast and mammalian cell lines.

Main Results:

The DRCR system successfully induced both intra- and extrachromosomal gene amplification.
Observed significant chromosome rearrangements, characteristic of gene amplification events.
Demonstrated scattered-type amplification patterns similar to those found in cancer cells.

Conclusions:

The developed DRCR system serves as a valuable model for studying oncogene and drug-resistance gene amplification.
This system can potentially enhance existing amplification strategies for pharmaceutical protein production in mammalian cells.