Binary recombinase systems for high-resolution conditional mutagenesis

Nucleic Acids Res. 2014 Apr;42(6):3894-907. doi: 10.1093/nar/gkt1361. Epub 2014 Jan 9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism
  • Cell Line
  • Genes, Reporter
  • HEK293 Cells
  • Humans
  • Integrases / metabolism*
  • Mice
  • Mutagenesis*
  • Neocortex / metabolism
  • Recombinases / metabolism*
  • Recombination, Genetic

Substances

  • Recombinases
  • Cre recombinase
  • Integrases