Nucleic Acids Research, Vol 24, Issue 23 4608-4613, Copyright © 1996 by Oxford University Press
B Sauer
Variant lox sites having an altered spacer region (heterospecific lox
sites) are not proficient for Cre-mediated recombination with the canonical
34 bp loxP site, but can recombine with each other. By placing different
heterospecific lox sites at different genomic locations, Cre can catalyze
independent DNA recombination events at multiple loci in the same cell
without concern that unwanted inter- locus recombination events will be
generated. Such heterospecific lox sites also allow Cre to specifically
target efficient integration of exogenous DNA to endogenous lox-like
sequences that naturally occur in the genome. Specific targeting occurs
only with a DNA vector carrying a heterospecific lox site in which the
spacer region has been redesigned to match the 'spacer' region of the
targeted chromosomal element. Moreover, in cells expressing a catalytically
active Cre recombinase, naturally occurring lox-like sequences can exhibit
almost 20% mitotic recombination. Thus, in the same cell, heterospecific
lox sites can be used independently at multiple loci for integration, for
deletion and for enhanced mitotic recombination, thereby increasing the
repertoire of genomic manipulations catalyzed by the Cre recombinase.
ARTICLES
Multiplex Cre/lox recombination permits selective site-specific DNA targeting to both a natural and an engineered site in the yeast genome
National Institutes of Health, National Institute of Diabetes, Digestive and Kidney Disease, Bethesda, MD 20892-1800, USA. sauerb@helix.nih.gov
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