Tokyo-Based Genomics Company Launches World’s First Cell-Free DNA Amplification Process

By T. Chakraborty, Ph.D.

DNA, the carrier of genetic information in cells, is the backbone of synthetic biology. Cells contain the required materials to amplify DNA molecules while making copies of them every time they split. For decades researchers have leveraged E. coli cells to amplify DNA for various uses ranging from the production of insulin to basic research. However, these cells have certain limitations.

Researchers have struggled over the years with DNA sequences that produce toxic proteins that cause harm to the E. coli, while it is extremely difficult to amplify GC-rich in the bacteria. OriCiro is a company pioneering in cell-free synthesis and amplification of genome-scale large DNA for gene therapy and synthetic biology. On September 30th, it announced the OriCiro Cell-Free Cloning System, the world’s first technology enabling cell-free amplification, which has taken the best features of E. coli and eliminated amplification of certain DNA sequences.

Seiji Hirasaki, CEO and Co-Founder of OriCiro Genomics, said, “Cell-free technology is one of the most important enabling technologies in synthetic biology because it can not only enhance the efficiency but also widen the scope of R&D. Our core technology is about cell-free assembly and amplification of genome-scale, large [<1 Mbp] DNA. We want to eliminate the need for E. coli and cell-based cloning for producing large DNA.”

Workings of the OriCiro Cell-Free System

The company, founded last year, focuses on genome propagation in E. coli, which is ten times faster. Dr. Masayuki Su’etsugu, OriCiro’s CSO and others, identified the essential proteins required for genome replication in E. coli, after which they constructed the entire process outside the cell. A single circular DNA with an oriC sequence is all that is required to begin DNA amplification. The OriCiro system can achieve 1010-fold amplification from a single DNA, which can be as large as 1Mbp, within 3 hours.

The Cell-Free Cloning System consists of the OriCiro Assembly Kit and OriCiro Amp Kit. The OriCiro Assembly kit is a single-step process that effortlessly assembles multiple DNA fragments by homologous overlapping ends. Their proprietary enzyme mix speeds up the annealing process of up to 50 DNA fragments in mere 30 minutes. The OriCiro Amp kit comprises a mix of 26 purified proteins that reconstructs the E. coli genome production in vitro. It requires several hours of isothermal incubation to yield a high concentration of pure, super-coiled DNA from circularized assembled molecules.

Dr. Masayuki Su’etsugu added, “OriCiro Cell-Free Cloning System removes the bottlenecks of conventional E. coli cloning and unleashes the potential of synthetic biology enabling efficient engineering of biological functions of microbes”.

Editor: Rajaneesh K. Gopinath, Ph.D.

Related Article: Codexis, Molecular Assemblies Enter Revolutionary DNA Synthesis Partnership

References

1. https://www.prnewswire.com/news-releases/oriciro-launches-worlds-first-cell-free-amplification-technology-for-large-circular-dna-301142005.html

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Author

Tulip Chakraborty