In vivo genetic ablation by Cre‐mediated expression of diphtheria toxin fragment A

A Ivanova, M Signore, N Caro, NDE Greene, AJ Copp… - genesis, 2005 - Wiley Online Library
genesis, 2005Wiley Online Library
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 …
Abstract
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. genesis 43:129–135, 2005. © 2005 Wiley‐Liss, Inc.
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