Nucleic Acids Research, Vol 25, Issue 4 868-872, Copyright © 1997 by Oxford University Press
K Araki, M Araki and K Yamamura
Site-directed DNA integration has been achieved by using a pair of mutant
lox sites, a right element (RE) mutant lox site and a left element (LE)
mutant lox site [Albertet al. (1995)Plant J., 7, 649-659], in mouse
embryonic stem (ES) cells. We established ES cell lines carrying a single
copy of the wild-type lox Por LE mutant lox site as a target and examined
the frequency of site-specific integration of a targeting vector carrying a
loxP or RE mutant lox site induced by Cre transient expression. Since our
targeting vector contains a complete neo gene, random integrants can form
colonies as in the case of a gene targeting event through homologous
recombination. With our system, the frequency of site-specific integration
via the mutant lox sites reached a maximum of 16%. In contrast, the
wild-type loxP sites yielded very low frequencies (<0.5%) of
site-specific integration events. This mutatedloxsystem will be useful for
'knock-in' integration of DNA in ES cells.
ARTICLES
Targeted integration of DNA using mutant lox sites in embryonic stem cells
Department of Developmental Genetics, Institute of Molecular Embryology and Genetics, Kumamoto University School of Medicine, Japan.
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