Eliminiating Any Regional Effect of a Knockout Removal of a Selectable Marker

To identify targeted events, an introduction of a positive selectable marker, usually neomycin, into the targeted locus is required. Recently, there has been an increasing concern regarding the repressor effect of the selectable marker cassette on the genes in the vicinity of its insertion. Therefore, removal of the marker from all targeted genes is advisable. This can most easily be performed by flanking (floxing[flanking with loxPI) the selectable marker cassette by loxP sites, which on introduction of the Cre recombinase will result in the removal of neomycin.

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