Conditional gene targeting, based on the site-specific recombination system such as Cre-loxP, plays a vital role in the study of gene functions and the generation of disease mouse models. It was always under consideration that there were problems in the Cre-loxP recombination system, such as illegal expression pattern of Cre transgene, variation of Cre recombination efficiency and toxicity of Cre recombinase, as well as the potential influences of genetic background, breeding strategy, experimental control and gene compensation. Oversights of these issues may have a profound influence on the accuracy of gene functional dissection and conditional gene targeting mice phenotypic interpretation. Accordingly, solutions should be adopted including delicate regulative control of temporal-spatial specific Cre expression, detailed detection of Cre recombination efficiency, reduction of Cre toxicity, simplification of mouse genetic background, optimization of breeding, setting up of proper control and combined conditional gene targeting.
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