Human genomic site-specific recombination catalyzed by coliphage HK022 integrase

It has been previously demonstrated that the wild type integrase (Int) protein of coliphage HK022 can catalyze site-specific recombination in human cells between attachment ( att) sites that were placed on extrachromosomal plasmids. In the present report it is shown that Int can catalyze the site-sp...

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Bibliographic Details
Published in:Journal of biotechnology 2008-03, Vol.134 (1), p.46-54
Main Authors: Harel-Levi, Gali, Goltsman, Janna, Tuby, Chen Nahum Josef Haim, Yagil, Ezra, Kolot, Mikhail
Format: Article
Language:English
Subjects:
Bacteriophage HK022 - enzymology
Biological and medical sciences
Biotechnology
Blotting, Southern
Cell Line
Coliphage HK022
DNA - genetics
DNA - metabolism
Escherichia coli
Fundamental and applied biological sciences. Psychology
Genome, Human - genetics
Human genome
Humans
Integrase
Integrases - metabolism
Models, Genetic
Plasmids - genetics
Recombination, Genetic - genetics
Site-specific recombination
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Summary:It has been previously demonstrated that the wild type integrase (Int) protein of coliphage HK022 can catalyze site-specific recombination in human cells between attachment ( att) sites that were placed on extrachromosomal plasmids. In the present report it is shown that Int can catalyze the site-specific recombination reactions in a human cell culture on the chromosomal level. These include integrative ( attP × attB) as well as excisive ( attL × attR) reactions each in two configurations. In the cis configuration both sites are on the same chromosome, in the trans configuration one site is on a chromosome and the other on an episome. The reactions in cis were observed without any selection force, using the green fluorescent protein (GFP) as a reporter. The reactions in trans could be detected only when a selection force was applied, using the hygromycin-resistant (Hyg R) phenotype as a selective marker. All reactions were catalyzed without the need to supply any of the accessory proteins that are required by Int in its Escherichia coli host. The versatility of the att sites may be an advantage in the utilization of Int to integrate plasmid DNA into the genome, followed by a partial exclusion of the integrated plasmid.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2008.01.002