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A Cre-Lox P Recombination Approach for the Detection of Cell Fusion In Vivo

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Published on: January 4, 2012

FLP and Cre recombinase function in Xenopus embryos.

D Werdien¹, G Peiler, G U Ryffel

¹Universitätsklinikum Essen, Institut für Zellbiologie (Tumorforschung), Hufelandstrasse 55, D-45122 Essen, Germany.

Nucleic Acids Research

|May 29, 2001

Summary

Researchers explored FLP and Cre recombinase systems in Xenopus embryos. FLP-mediated gene activation was controllable, while Cre showed higher efficiency, enabling new transgenic Xenopus research.

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Area of Science:

Developmental Biology
Molecular Biology
Genetics

Background:

Site-specific DNA recombinases FLP and Cre are widely used in various organisms.
Their application in the Xenopus system, a crucial model for vertebrate development, remained uninvestigated.

Purpose of the Study:

To evaluate the efficacy and applicability of FLP and Cre recombinase systems in Xenopus embryos.
To develop novel inducible and dual-reporter gene expression systems for Xenopus.

Main Methods:

Injection of mRNA encoding FLP or Cre recombinase into Xenopus embryos.
Utilized reporter constructs with FLP recombination target (FRT) or loxP sites flanking reporter genes (GFP, lacZ).
Developed an inducible FLP system using a fusion protein dependent on 4-hydroxytamoxifen and dual-reporter systems.

Main Results:

FLP recombinase successfully mediated excision of FRT-flanked sequences, activating downstream gene expression (lacZ).
An inducible FLP system allowed temporal control of gene activation in Xenopus embryos.
Cre recombinase demonstrated high recombination efficiency in early Xenopus embryos, preceding zygotic transcription.
Cre recombinase was found to be more effective than FLP in Xenopus for a given reporter quantity.
Novel FLP-inducible dual-reporter systems were established for simultaneous analysis of two fluorescent proteins.

Conclusions:

Both FLP and Cre recombinase systems are applicable and functional in Xenopus embryos.
Cre recombinase exhibits superior efficiency in Xenopus recombination assays compared to FLP.
The developed inducible and dual-reporter systems offer powerful tools for genetic manipulation and lineage tracing in transgenic Xenopus.
These findings pave the way for advanced cell-lineage studies in Xenopus using recombinase-mediated DNA rearrangement.