Binary recombinase systems for high-resolution conditional mutagenesis
Nucleic Acids Research, 2014•academic.oup.com
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 …
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 …
Abstract
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.
Oxford University Press