Binary recombinase systems for high-resolution conditional mutagenesis.

Hermann, Mario; Stillhard, Patrick; Wildner, Hendrik; Seruggia, Davide; Kapp, Viktor; Sánchez-Iranzo, Héctor; Mercader Huber, Nadia; Montoliu, Lluís; Zeilhofer, Hanns Ulrich; Pelczar, Pawel (2014). Binary recombinase systems for high-resolution conditional mutagenesis. Nucleic acids research, 42(6), pp. 3894-3907. Information Retrieval Ltd. 10.1093/nar/gkt1361

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Conditional mutagenesis using Cre recombinase expressed from tissue specific promoters facilitates analyses of gene function and cell lineage tracing. Here, we describe two novel dual-promoter-driven conditional mutagenesis systems designed for greater accuracy and optimal efficiency of recombination. Co-Driver employs a recombinase cascade of Dre and Dre-respondent Cre, which processes loxP-flanked alleles only when both recombinases are expressed in a predetermined temporal sequence. This unique property makes Co-Driver ideal for sequential lineage tracing studies aimed at unraveling the relationships between cellular precursors and mature cell types. Co-InCre was designed for highly efficient intersectional conditional transgenesis. It relies on highly active trans-splicing inteins and promoters with simultaneous transcriptional activity to reconstitute Cre recombinase from two inactive precursor fragments. By generating native Cre, Co-InCre attains recombination rates that exceed all other binary SSR systems evaluated in this study. Both Co-Driver and Co-InCre significantly extend the utility of existing Cre-responsive alleles.

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