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Cell-free synthesis and amplification of genome-scale large DNA

01 SUMMARY

Cell-free assembly and amplification aimed at replacing cell-based cloning

OriCiro offers ground breaking, cell-free technologies of large DNA assembly and amplification that can effectively replace the conventional E. coli cloning process.

02 ASSEMBLY

Hyper assembly of multiple DNA fragments

  • One-step isothermal reaction, cell-free, easy-to-handle
  • Seamless assembly via overlapping homologous ends (25 - 60 bp)
  • Simultaneous assembly of up to 50 fragments
  • Assembly of large DNA fragments up to 200 kb

03 AMPLIFICATION

In-vitro reconstitution of chromosome replication cycle

  • Whole replication cycle of the E. coli chromosome is reconstituted in vitro using purified proteins.
  • The replication cycle repeats autonomously and continuously allowing to "exponential" amplification of circular DNA.
  • Requirements for the template DNA are "circular configuration" and "oriC sequence (300 bp)."

Link to the relevant scientific publication

Cell-free hyper amplification of large DNA

  • One-step isothermal reaction, cell-free, easy-to-handle
  • Up to 100 ng/µl supercoiled DNA produced within 3 hours at 30ºC even from a single DNA molecule (for 10 kbp DNA)
  • Extremely high fidelity
  • Applicable to a wide range of DNA size from 1 kbp to 1 Mbp
  • Applicable to any sequence including GC-rich and repeat sequences

04 COMBINATION

Efficient cell-free production of circular DNA from multiple DNA fragments

  • Apply the assembled products directly to the amplification reaction.
  • Circular DNA (complete assembled products) can be amplified selectively from a single molecule level.
  • No need of purification process after assembly as unnecessary intermediates (linear DNA) are not amplified.

05 STRENGTH

Distinctive advantages of our amplification technology

OriCiro technology has a number of advantages over existing approaches making our technology very unique and competitive.
PCR E. coli cloning OriCiro Technology
DNA size +
(10 kbp)
++
(300 kbp)
+++
(1 Mbp)
Operation Thermocycler Cumbersome process
Several days requiring techniques
Very simple process
Several hours of
isothermal incubation
Biosafety Cell-free Recombinant DNA experiment Cell-free
Fidelity +
(10-4~10-6 error/bp)
+++
(10-10 error/bp)
++
(10-8 error/bp)
Sequence
applicability
Not applicable to GC
rich and repeat sequences
Not applicable to sequences
that cause cell toxicity
Applicable to any sequence
Product Linear DNA Circular DNA Circular DNA

※Technology under development to match E. coli cloning.

06 PRODUCTS & SERVICES

  • 01

    Reagent kits

    Reagent kits for assembly and amplification of large DNA for academia research

  • 02

    Co-development

    Co-development arrangements with partners from the pharma, diagnostics, agriculture and industrial biotech companies

  • 03

    Technology licensing

    Technology licensing to CDMOs and gene synthesis companies for using our proprietary technologies

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