Efficient Flp-Int HK022 dual RMCE in mammalian cells

Eugenia Voziyanova, Natalia Malchin, Rachelle P. Anderson, Ezra Yagil, Mikhail Kolot, Yuri Voziyanov*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Recombinase-mediated cassette exchange, or RMCE, is a clean approach of gene delivery into a desired chromosomal location, as it is able to insert only the required sequences, leaving behind the unwanted ones. RMCE can be mediated by a single site-specific DNA recombinase or by two recombinases with different target specificities (dual RMCE). Recently, using the Flp-Cre recombinase pair, dual RMCE proved to be efficient, provided the relative ratio of the enzymes during the reaction is optimal. In the present report, we analyzed how the efficiency of dual RMCE mediated by the Flp-Int (HK022) pair depends on the variable input of the recombinases-the amount of the recombinase expression vectors added at transfection-and on the order of the addition of these vectors: sequential or simultaneous. We found that both in the sequential and the simultaneous modes, the efficiency of dual RMCE was critically dependent on the absolute and the relative concentrations of the Flp and Int expression vectors. Under optimal conditions, the efficiency of 'simultaneous' dual RMCE reached ∼12% of the transfected cells. Our results underline the importance of fine-tuning the reaction conditions for achieving the highest levels of dual RMCE.

Original languageEnglish
Pages (from-to)e125
JournalNucleic Acids Research
Issue number12
StatePublished - Jul 2013


FundersFunder number
German Israeli Foundation for Scientific Research and Development1062-62.3/2008
National Institutes of Health
National Institute of General Medical SciencesR01GM085848
Israel Science Foundation702/11


    Dive into the research topics of 'Efficient Flp-Int HK022 dual RMCE in mammalian cells'. Together they form a unique fingerprint.

    Cite this