Gibson Deletion: A novel application of isothermal in vitro recombination

Swara Kalva, Jef D. Boeke, Paolo Mita

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Background: Recombinant DNA technology is today a fundamental tool for virtually all biological research fields. Among the many techniques available for the construction of a "custom DNA" molecule, the isothermal in vitro assembly, or Gibson assembly, allows for an efficient, one-step, scarless recombination-based assembly. Results: Here, we apply and characterize the use of Gibson assembly for the deletion of DNA sequences around a DNA cut. This method, that we named "Gibson Deletion", can be used to easily substitute or delete one or more restriction sites within a DNA molecule. We show that Gibson Deletion is a viable method to delete up to 100 nucleotides from the DNA ends of a cleavage site. In addition, we found that Gibson Deletion can be performed using single strand DNA with the same efficiency as using double strand DNA molecules. Conclusions: Gibson Deletion is a novel, easy and convenient application of isothermal in vitro assembly, that performs with high efficiency and can be implemented for a broad range of applications.

    Original languageEnglish (US)
    Article number2
    JournalBiological Procedures Online
    Volume20
    Issue number1
    DOIs
    StatePublished - Jan 19 2018

    Keywords

    • Cloning
    • Deletion
    • Gibson assembly
    • Isothermal in vitro assembly
    • Restriction site

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

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