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An improved mouse line for Cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus

An improved mouse line for Cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus
An improved mouse line for Cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus
The means to specifically ablate cells inside of a living organism have recently been improved and facilitated by stable mouse lines, carrying conditional expression constructs for diphtheria toxin (DT) or diphtheria toxin receptor, that could be activated upon Cre-mediated recombination or the application of diphtheria toxin, respectively. We have lately described the R26:lacZ/DT-A line (Brockschnieder et al., 2004, Mol Cell Biol 24:7636-7642) in which a loxP-conditional DTA allele was introduced into the ubiquitously expressed Rosa26 locus. This strain allowed the ablation of a wide spectrum of cell types by crossing it to tissue specific Cre lines. Unexpectedly, homozygous (but not heterozygous) animals of the R26:lacZ/DT-A line developed some degenerative abnormalities in a variety of tissues. The defects were most probably caused by leaky expression of small amounts of toxin from the unrecombined lacZ(flox)DT-A cassette. Here we show that insertion of an additional transcriptional regulatory sequence (bovine growth hormone polyadenylation signal, bpA) following the lacZ open reading frame prevented the formation of any defects in homozygous mice. The modification did not affect the functionality of the lacZ(flox)DTA allele, as exemplified by the complete ablation of oligodendrocytes upon Cre-mediated recombination. The novel R26:lacZbpA(flox)DTA line is expected to greatly facilitate the reliable generation of cell type ablated mice.
affect, peptide fragments, integrases, suicide, abnormalities, biology, mice, diphtheria toxin, metabolism, mutagenesis, lac operon, protein, inbred c57bl, gene expression regulation, research support, research, growth hormone, homozygote, genes, transgenic, oligodendroglia, expression, growth, insertional, animals, genetics, proteins
1526-954X
322-327
Brockschnieder, D.
766bdfda-70f4-4c9d-82e4-96ab38497161
Pechmann, Y.
fdb0c4e9-28f1-4332-9588-fb68672c8822
Sonnenberg-Riethmacher, E.
6da24b56-bcab-43c2-8da3-01c99f274f1d
Riethmacher, D.
1a0a0c2e-e94d-4d0a-a890-90107a2545bc
Brockschnieder, D.
766bdfda-70f4-4c9d-82e4-96ab38497161
Pechmann, Y.
fdb0c4e9-28f1-4332-9588-fb68672c8822
Sonnenberg-Riethmacher, E.
6da24b56-bcab-43c2-8da3-01c99f274f1d
Riethmacher, D.
1a0a0c2e-e94d-4d0a-a890-90107a2545bc

Brockschnieder, D., Pechmann, Y., Sonnenberg-Riethmacher, E. and Riethmacher, D. (2006) An improved mouse line for Cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus. Genesis, 44 (7), 322-327. (doi:10.1002/dvg.20218).

Record type: Article

Abstract

The means to specifically ablate cells inside of a living organism have recently been improved and facilitated by stable mouse lines, carrying conditional expression constructs for diphtheria toxin (DT) or diphtheria toxin receptor, that could be activated upon Cre-mediated recombination or the application of diphtheria toxin, respectively. We have lately described the R26:lacZ/DT-A line (Brockschnieder et al., 2004, Mol Cell Biol 24:7636-7642) in which a loxP-conditional DTA allele was introduced into the ubiquitously expressed Rosa26 locus. This strain allowed the ablation of a wide spectrum of cell types by crossing it to tissue specific Cre lines. Unexpectedly, homozygous (but not heterozygous) animals of the R26:lacZ/DT-A line developed some degenerative abnormalities in a variety of tissues. The defects were most probably caused by leaky expression of small amounts of toxin from the unrecombined lacZ(flox)DT-A cassette. Here we show that insertion of an additional transcriptional regulatory sequence (bovine growth hormone polyadenylation signal, bpA) following the lacZ open reading frame prevented the formation of any defects in homozygous mice. The modification did not affect the functionality of the lacZ(flox)DTA allele, as exemplified by the complete ablation of oligodendrocytes upon Cre-mediated recombination. The novel R26:lacZbpA(flox)DTA line is expected to greatly facilitate the reliable generation of cell type ablated mice.

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More information

Published date: 21 June 2006
Keywords: affect, peptide fragments, integrases, suicide, abnormalities, biology, mice, diphtheria toxin, metabolism, mutagenesis, lac operon, protein, inbred c57bl, gene expression regulation, research support, research, growth hormone, homozygote, genes, transgenic, oligodendroglia, expression, growth, insertional, animals, genetics, proteins

Identifiers

Local EPrints ID: 59525
URI: https://eprints.soton.ac.uk/id/eprint/59525
ISSN: 1526-954X
PURE UUID: d9c26ddf-59cb-456c-97dc-93f3f92b45de

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Date deposited: 03 Sep 2008
Last modified: 13 Mar 2019 20:30

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