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

Gene Therapy
Mikhail KolotEzra Yagil

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.

References

Jan 20, 1985·Journal of Molecular Biology·C E BauerR I Gumport
Mar 11, 1986·Nucleic Acids Research·R H HoessK Abremski
Jun 14, 1967·Journal of Molecular Biology·B Hirt
Dec 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·W Ross, A Landy
Dec 1, 1994·Molecular & General Genetics : MGG·M Kolot, E Yagil
Oct 30, 1998·American Journal of Human Genetics·V ShotelersukW A Gahl
Jan 15, 1999·Human Molecular Genetics·N SandovalD Schindler
Feb 26, 2000·Annual Review of Genetics·R A WeisbergJ W Little
Oct 13, 2001·Advances in Genetics·C Hansis, J Grifo
Nov 2, 2001·Nature Biotechnology·F Buchholz, A F Stewart
Aug 12, 2003·Biotechnology and Bioengineering·Mikhail KolotEzra Yagil
Jan 16, 2004·Developmental Cell·Catherine S Branda, Susan M Dymecki
Apr 28, 2004·Plasmid·Nadège PhilippeDominique Schneider
Mar 3, 2005·Nucleic Acids Research·Gilda CobellisRiccardo Cortese
Jun 15, 2007·Applied and Environmental Microbiology·Susan R Steyert, Silvia A Pineiro
Jun 30, 2007·Science·Indrani SarkarFrank Buchholz
Oct 2, 2007·Current Opinion in Biotechnology·Dagmar WirthHansjörg Hauser
Mar 1, 2008·Science·Jemeen SreedharanChristopher E Shaw
Sep 25, 2008·Nature Reviews. Molecular Cell Biology·Martin F Lavin
Mar 10, 2009·Gene·Natalia MalchinMikhail Kolot
Jan 19, 2011·Journal of Molecular Biology·Soeren TuranJuergen Bode
Mar 29, 2011·Molecular Genetics and Genomics : MGG·Natalia MalchinMikhail Kolot
Jan 1, 2013·Nucleic Acids Research·Josephine Abi-GhanemM Teresa Pisabarro
May 1, 2013·Nucleic Acids Research·Eugenia VoziyanovaYuri Voziyanov

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Citations

Apr 27, 2016·Journal of Molecular Biology·Bryce L Lunt, Graham F Hatfull
Sep 1, 2018·Plant Cell Reports·Peter G KrothAngela Falciatore
May 28, 2020·ACS Synthetic Biology·Takahito MukaiMasayuki Su'etsugu

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