Binary recombinase systems for high-resolution conditional mutagenesis

M Hermann, P Stillhard, H Wildner… - Nucleic Acids …, 2014 - academic.oup.com
M Hermann, P Stillhard, H Wildner, D Seruggia, V Kapp, H Sanchez-Iranzo, N Mercader…
Nucleic Acids Research, 2014academic.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 …
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