In vivo genetic ablation by Cre-mediated expression of diphtheria toxin fragment A.
We generated a ROSA26-eGFP-DTA mouse line by introducing an eGFP-DTA (enhanced green fluorescent protein -- diphtheria toxin fragment A) cassette into the ROSA26 locus by homologous recombination in ES cells. This mouse expresses eGFP ubiquitously, but DTA expression is prevented by the presence of eGFP, a Neo cassette, and a strong transcriptional stop sequence. Mice carrying this construct are normal and fertile, indicating the absence of DTA expression. However, upon Cre-mediated excision of the floxed region DTA expression is activated, resulting in the specific ablation of Cre-expressing cells. As an example of this approach, we ablated Nkx2.5 and Wnt1-expressing cells by using the Nkx2.5-Cre and Wnt1-Cre mouse lines, respectively. We observed loss of the precise tissues in which Nkx2.5 and Wnt1 are expressed. Apart from being a general GFP reporter, the ROSA26-GFP-DTA mouse line should provide a useful resource for genetic ablation of specific groups of cells.
Ivanova
Anna
A
Neural Development Unit, Institute of Child Health, University College London, United Kingdom.
Signore
Massimo
M
Caro
Nadia
N
Greene
Nicholas D E
ND
Copp
Andrew J
AJ
Martinez-Barbera
Juan Pedro
JP
D000818:Animals; D001921:Brain / Q000196:embryology; D016923:Cell Death; D004167:Diphtheria Toxin / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006321:Heart / Q000196:embryology; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006824:Hybridization, Genetic; D017403:In Situ Hybridization; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D051155:Wnt1 Protein / Q000235:genetics / Q000378:metabolism
D004167:Diphtheria Toxin; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C484469:Nkx2-5 protein, mouse; D010446:Peptide Fragments; D014157:Transcription Factors; D051155:Wnt1 Protein; C495246:Wnt1 protein, mouse; C029795:diphtheria toxin fragment A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins