A global double‐fluorescent Cre reporter mouse

The Cre/loxP system has been used extensively for conditional mutagenesis in mice. Reporters of Cre activity are important for defining the spatial and temporal extent of Cre‐mediated recombination. Here we describe mT/mG, a double‐fluorescent Cre reporter mouse that expresses membrane‐targeted tandem dimer Tomato (mT) prior to Cre‐mediated excision and membrane‐targeted green fluorescent protein (mG) after excision. We show that reporter expression is nearly ubiquitous, allowing visualization of fluorescent markers in live and fixed samples of all tissues examined. We further demonstrate that mG labeling is Cre‐dependent, complementary to mT at single cell resolution, and distinguishable by fluorescence‐activated cell sorting. Both membrane‐targeted markers outline cell morphology, highlight membrane structures, and permit visualization of fine cellular processes. In addition to serving as a global Cre reporter, the mT/mG mouse may also be used as a tool for lineage tracing, transplantation studies, and analysis of cell morphology in vivo. genesis 45:593–605, 2007. © 2007 Wiley‐Liss, Inc.

[1]  A. Nagy,et al.  Creation and use of a cre recombinase transgenic database. , 2009, Methods in molecular biology.

[2]  L. Luo,et al.  Modeling sporadic loss of heterozygosity in mice by using mosaic analysis with double markers (MADM) , 2007, Proceedings of the National Academy of Sciences.

[3]  H. Fehling,et al.  Faithful activation of an extra‐bright red fluorescent protein in “knock‐in” Cre‐reporter mice ideally suited for lineage tracing studies , 2007, European journal of immunology.

[4]  V. Raman,et al.  Noninvasive optical tracking of red fluorescent protein-expressing cancer cells in a model of metastatic breast cancer. , 2006, Neoplasia.

[5]  Nathan C Shaner,et al.  A guide to choosing fluorescent proteins , 2005, Nature Methods.

[6]  Min Han,et al.  Efficient Transposition of the piggyBac (PB) Transposon in Mammalian Cells and Mice , 2005, Cell.

[7]  J. Long,et al.  Genetic and spectrally distinct in vivo imaging: embryonic stem cells and mice with widespread expression of a monomeric red fluorescent protein , 2005, BMC biotechnology.

[8]  Min Han,et al.  Ubiquitous expression of mRFP1 in transgenic mice , 2005, Genesis.

[9]  L. Luo,et al.  Mosaic Analysis with Double Markers in Mice , 2005, Cell.

[10]  Min Han,et al.  Ubiquitous expression of mRFP1 in transgenic mice , 2005, Genesis.

[11]  M. Gertsenstein,et al.  Mouse in red: Red fluorescent protein expression in mouse ES cells, embryos, and adult animals , 2004, Genesis.

[12]  R. Tsien,et al.  Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein , 2004, Nature Biotechnology.

[13]  Vincenzo De Paola,et al.  AMPA receptors regulate dynamic equilibrium of presynaptic terminals in mature hippocampal networks , 2003, Nature Neuroscience.

[14]  N. Copeland,et al.  A highly efficient recombineering-based method for generating conditional knockout mutations. , 2003, Genome research.

[15]  Y. Jan,et al.  Progenitor cell maintenance requires numb and numblike during mouse neurogenesis , 2002, Nature.

[16]  Andrew P McMahon,et al.  Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. , 2002, Developmental biology.

[17]  Shi Tang,et al.  A cre/loxP‐deleter transgenic line in mouse strain 129S1/SvImJ , 2002, Genesis.

[18]  Caiying Guo,et al.  A cre recombinase transgene with mosaic, widespread tamoxifen‐inducible action , 2002, Genesis.

[19]  B. Glick,et al.  Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed) , 2002, Nature Biotechnology.

[20]  Shankar Srinivas,et al.  Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus , 2001, BMC Developmental Biology.

[21]  Y Fujiwara,et al.  Activation of EGFP expression by Cre-mediated excision in a new ROSA26 reporter mouse strain. , 2001, Blood.

[22]  Caiying Guo,et al.  Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon cre‐mediated excision , 2000, Genesis.

[23]  R Y Tsien,et al.  Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Mcconnell,et al.  Targeting of cre to the Foxg1 (BF-1) locus mediates loxP recombination in the telencephalon and other developing head structures. , 2000, Developmental biology.

[25]  Andras Nagy,et al.  Cre recombinase: The universal reagent for genome tailoring , 2000, Genesis.

[26]  S. Lukyanov,et al.  Fluorescent proteins from nonbioluminescent Anthozoa species , 1999, Nature Biotechnology.

[27]  C. Lobe,et al.  Z/AP, a double reporter for cre-mediated recombination. , 1999, Developmental biology.

[28]  Philippe Soriano.  Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[29]  P. Caroni,et al.  The motility-associated proteins GAP-43, MARCKS, and CAP-23 share unique targeting and surface activity-inducing properties. , 1997, Experimental cell research.

[30]  K. Rajewsky,et al.  Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. , 1994, Science.

[31]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[32]  B. Sauer.  Manipulation of transgenes by site-specific recombination: use of Cre recombinase. , 1993, Methods in enzymology.

[33]  Yamamura Ken-ichi,et al.  Efficient selection for high-expression transfectants with a novel eukaryotic vector , 1991 .

[34]  H. Niwa,et al.  Efficient selection for high-expression transfectants with a novel eukaryotic vector. , 1991, Gene.

[35]  R. McKay,et al.  CNS stem cells express a new class of intermediate filament protein , 1990, Cell.

[36]  P. Greengard,et al.  Molecular cloning, characterization, and expression of a cDNA encoding the "80- to 87-kDa" myristoylated alanine-rich C kinase substrate: a major cellular substrate for protein kinase C. , 1989, Proceedings of the National Academy of Sciences of the United States of America.