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  • Original Article
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Site promiscuity of coliphage HK022 integrase as a tool for gene therapy

Gene Therapy volume 22, pages 521–527 (2015)Cite this article

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A Corrigendum to this article was published on 02 July 2015

This article has been updated

Abstract

The integrase (Int) encoded by the lambdoid coliphage HK022 targets in its host chromosome a 21 base pair (bp) recombination site termed attB or BOB'. attB comprises two 7 bp partially inverted (palindromic) Int-binding sites of 7 bp each termed B and B'. B and B' flank a central 7 bp crossover site or ‘overlap’ (O). We show that replacing O with a random 7 bp sequence supports Int-mediated site-specific recombination as long as the cognate and larger phage recombination site attP features an identical O sequence. This promiscuity allowed us to identify on the human genome several native active secondary attB sites (‘attB’) with random overlaps that flank human deleterious mutations, raising the prospect of using such sites to cure the ‘attB’-flanked mutations by Int-catalyzed RMCE (recombinase-mediated cassette exchange) reactions. An analysis of such active and inactive ‘attB’s suggested a minimal 14–15 bp attB consensus sequence (instead of the 21 bp) with a reduced 3 bp palindrome.

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Change history

  • 02 July 2014

    This article has been corrected since advance online publication and a corrigendum is also printed in this issue

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Acknowledgements

Jury Voziyanov kindly supplied and guided us with the Target Finder program and Gabriel Kaufmann assisted us with helpful comments. Research was supported by GIF, the German Israeli Foundation for Scientific Research and development (Grant 1062/2008 to EY and MK), and the Israel Science Foundation (Grant 702/11 to EY and MK).

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  1. M Kolot and N Malchin: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Tel-Aviv University, Tel-Aviv, Israel

    M Kolot, N Malchin, A Elias, N Gritsenko & E Yagil

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Correspondence to M Kolot or E Yagil.

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Kolot, M., Malchin, N., Elias, A. et al. Site promiscuity of coliphage HK022 integrase as a tool for gene therapy. Gene Ther 22, 521–527 (2015). https://doi.org/10.1038/gt.2015.9

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