The IRG mouse: a two-color fluorescent reporter for assessing Cre-mediated recombination and imaging complex cellular relationships in situ.
The Cre-loxP system is widely used for making conditional alterations to the mouse genome. Cre-mediated recombination is frequently monitored using reporter lines in which Cre expression activates a reporter gene driven by a ubiquitous promoter. Given the distinct advantages of fluorescent reporters, we developed a transgenic reporter line, termed IRG, in which DsRed-Express, a red fluorescent protein (RFP) is expressed ubiquitously prior to Cre-mediated recombination and an enhanced green fluorescent protein (EGFP) following recombination. Besides their utility for monitoring Cre-mediated recombination, we show that in IRG mice red and green native fluorescence can be imaged simultaneously in thick tissue sections by confocal microscopy allowing for complex reconstructions to be created that are suitable for analysis of neuronal morphologies as well as neurovascular interactions in brain. IRG mice should provide a versatile tool for analyzing complex cellular relationships in both neural and nonneural tissues.
Research and Development, James J. Peters Department of Veterans Affairs Medical Center, Bronx, New York 10468, USA.
Rita
R
De Gasperi
Anne B
AB
Rocher
Miguel A Gama
MA
Sosa
Susan L
SL
Wearne
Gissel M
GM
Perez
Victor L
VL
Friedrich
Patrick R
PR
Hof
Gregory A
GA
Elder
D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D003433:Crosses, Genetic; D003952:Diagnostic Imaging; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism*; D017930:Genes, Reporter; D005819:Genetic Markers / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D007381:Intermediate Filament Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000235:genetics; D064231:Nestin; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014018:Tissue Distribution; D019076:Transgenes; D000097573:Red Fluorescent Protein
D005456:Fluorescent Dyes; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D019426:Integrases; D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; D000097573:Red Fluorescent Protein; C577485:Nes protein, mouse; C485184:enhanced green fluorescent protein; C045073:Cre recombinase