![]() The floxed and trapped alleles can typically be generated using a single construct bearing a gene-trap cassette doubly flanked by LoxP and FRT sites, and can be used independently to achieve conditional and constitutive gene KO, respectively. This method involves two alleles of a target gene: a standard floxed allele, and a multi-functional allele bearing an FRT-flanked gene-trap cassette, which inactivates the target gene while reporting its expression with green fluorescent protein (GFP) the trapped allele is thus a null and GFP reporter by default, but is convertible into a wild-type allele. We describe a simple method we have named LOFT, which is capable of reversible cKO and free of the limitations associated with existing techniques. ![]() A number of methods have been developed to overcome this, but each method has its own limitations. However, a major limitation of this method is that gene KO is not reversible. Conditional gene knockout (cKO) mediated by the Cre/LoxP system is indispensable for exploring gene functions in mice.
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