2012
DOI: 10.1073/pnas.1211816109
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Inducible genetic system for the axolotl

Abstract: Transgenesis promises a powerful means for assessing gene function during amphibian limb regeneration. This approach is complicated, however, by the need for embryonic appendage development to proceed unimpeded despite the genetic alterations one wishes to test later in the context of regeneration. Achieving conditional gene regulation in this amphibian has not proved to be as straightforward as in many other systems. In this report we describe a unique method for obtaining temporal control over exogenous gene… Show more

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Cited by 41 publications
(41 citation statements)
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“…Hence, we injected single-cell axolotl embryos with this plasmid to generate transgenic F 0 animals. Though embryos were injected when they were single cells, transgenics show mosaic expression of transgenes (Monaghan and Maden, 2012;Sobkow et al, 2006;Whited et al, 2012) that is likely to be due to integration of the transgenic element into the genome after cell division has commenced; we observed a similar phenomenon. EGFP fluorescence was evident in embryos within 5-6 days post-injection.…”
Section: Vascular Endothelial Cell-specific Retroviral Infectionssupporting
confidence: 60%
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“…Hence, we injected single-cell axolotl embryos with this plasmid to generate transgenic F 0 animals. Though embryos were injected when they were single cells, transgenics show mosaic expression of transgenes (Monaghan and Maden, 2012;Sobkow et al, 2006;Whited et al, 2012) that is likely to be due to integration of the transgenic element into the genome after cell division has commenced; we observed a similar phenomenon. EGFP fluorescence was evident in embryos within 5-6 days post-injection.…”
Section: Vascular Endothelial Cell-specific Retroviral Infectionssupporting
confidence: 60%
“…In the absence of Cre, cells with the reporter only express EGFP. When co-transfected with a plasmid encoding a constitutively active Cre, AL1 cells and limb blastemas express tdTomato because the floxed EGFP-stop cassette has been eliminated through recombination at the LoxP sites (Whited et al, 2012). Al1 cells transfected with the Cre reporter plasmid were infected with a VSV-pseudotyped QCAG-Cre virus (titer 10 8 ) 3 days later.…”
Section: Research Articlementioning
confidence: 99%
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“…Although off-target mutations can be selected against by outcrossing mutant lines for several generations, the time frame required by this approach is not practical for experiments with an organism with a long generation time. Thus, phenotypes should be confirmed in mutants created with sgRNAs directed against different targets within the same gene, and, when possible, rescue experiments should be conducted using recent innovations in exogenous gene delivery and inducible gene expression in the axolotl (Khattak et al, 2013b;Whited et al, 2012Whited et al, , 2013). …”
Section: Discussionmentioning
confidence: 99%
“…This suggests that adult regeneration may be an acquired trait in axolotls that might be inducible in mammals if the required genetic pathways are mapped. Recent advances in production of transgenic axolotls, [6][7][8][9][10] complete mapping of the axolotl transcriptome, 11 and production of gene expression arrays [12][13][14][15][16] finally allow molecular mapping of regeneration pathways. Additionally, given the extensive conservation of synteny between axolotls and humans, this amphibian may be a powerful genetic model to study hematopoietic cell function in scar-free wound healing and tissue regeneration.…”
Section: Introductionmentioning
confidence: 99%