In vivo analysis of mRNA stability using the Tet-Off system in the chicken embryo

Hilgers, Valérie; Pourquié, Olivier; Dubrulle, Julien

doi:10.1016/j.ydbio.2005.05.021

Cited by 38 publications

(43 citation statements)

References 60 publications

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“…Such polymorphisms may have functional effects on mRNA conformation or stability, and hence gene expression level. 50 However, we observed no differences in mRNA abundance between clinical groups or haplotypes. Conversely, miRNAs interfere with translation, so their regulatory effect will only be detectable at the protein level.…”

Section: Discussioncontrasting

confidence: 54%

A region of 35 kb containing the trace amine associate receptor 6 (TAAR6) gene is associated with schizophrenia in the Irish study of high-density schizophrenia families

et al. 2007

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The TAAR6 gene has been previously associated with schizophrenia in 192 pedigrees of European and African ancestry. To replicate these findings we performed an association study of TAAR6 in 265 pedigrees of the Irish Study of High-Density Schizophrenia Families (ISHDSF). Of the 24 genotyped single-nucleotide polymorphisms only rs12189813 and rs9389011 provided single-marker evidence for association (0.0094pPp0.03). Two-marker haplotypes (rs7772821 and rs12189813; 0.0071pPp0.0023) and four-marker haplotypes (rs8192622, rs7772821, rs12189813 and rs9389011; 0.0047pPp0.018) gave strongest evidence for association. The associated high-risk (HR) haplotype in the ISHDSF is defined by the major alleles at rs7772821 and rs12189813 (0.00097pPp0.023). The associated HR remains positive in a case only test of association by Operational Criteria score analysis in which significant association was observed only with the highest threshold for delusions (P < 0.009). When analysis was restricted to affected individuals under the core schizophrenia (D2) diagnosis, the observed associations for HR were most significant in the highest threshold for delusions (P < 0.004) and hallucinations (P < 0.0004), supporting the family-based association with schizophrenia.

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Section: Discussioncontrasting

confidence: 54%

A region of 35 kb containing the trace amine associate receptor 6 (TAAR6) gene is associated with schizophrenia in the Irish study of high-density schizophrenia families

et al. 2007

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“…What is more, this technique can be used to deliver plasmids that carry the gene of interest under a constitutive, cell-type specific or inducible promoter/enhancer, or plasmids designed to study promoters and enhancers by driving the expression of reporter genes ( [100-109]; and others). For example, Hilgers et al combined electroporation with the tetracycline-dependent inducible Tet-Off system to study the effects of the 3'UTR in mRNA stability in the embryonic chick [110], whereas Watanabe et al used electroporation of Tet-On and Tet-Off constructs to elucidate previously unknown roles of certain genes during chick somitogenesis [111]. …”

Section: Reviewmentioning

confidence: 99%

Rediscovering the chick embryo as a model to study retinal development

Vergara

Canto‐Soler

2012

Neural Dev

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The embryonic chick occupies a privileged place among animal models used in developmental studies. Its rapid development and accessibility for visualization and experimental manipulation are just some of the characteristics that have made it a vertebrate model of choice for more than two millennia. Until a few years ago, the inability to perform genetic manipulations constituted a major drawback of this system. However, the completion of the chicken genome project and the development of techniques to manipulate gene expression have allowed this classic animal model to enter the molecular age. Such techniques, combined with the embryological manipulations that this system is well known for, provide a unique toolkit to study the genetic basis of neural development. A major advantage of these approaches is that they permit targeted gene misexpression with extremely high spatiotemporal resolution and over a large range of developmental stages, allowing functional analysis at a level, speed and ease that is difficult to achieve in other systems. This article provides a general overview of the chick as a developmental model focusing more specifically on its application to the study of eye development. Special emphasis is given to the state of the art of the techniques that have made gene gain- and loss-of-function studies in this model a reality. In addition, we discuss some methodological considerations derived from our own experience that we believe will be beneficial to researchers working with this system.

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“…There are a very few assays to test RNA stability in vivo and they have been developed for specific circumstances(Hilgers, Pourquie et al, 2005;Lee, Bradley et al, 1998); we have not developed such an assay for the CRPV viral oncogene RNAs. Therefore, we used the Zuker M-Fold program, version 3.5, (Zuker, 2003) to check predicted stabilities of wild type and codon-modified genes, both E6 and E7.…”

Section: Discussionmentioning

confidence: 99%

Synonymous codon changes in the oncogenes of the cottontail rabbit papillomavirus lead to increased oncogenicity and immunogenicity of the virus

Cladel

Budgeon

et al. 2013

Virology

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Papillomaviruses use rare codons with respect to the host. The reasons for this are incompletely understood but among the hypotheses is the concept that rare codons result in low protein production and this allows the virus to escape immune surveillance. We changed rare codons in the oncogenes E6 and E7 of the cottontail rabbit papillomavirus to make them more mammalian-like and tested the mutant genomes in our in vivo animal model. While the amino acid sequences of the proteins remained unchanged, the oncogenic potential of some of the altered genomes increased dramatically. In addition, increased immunogenicity, as measured by spontaneous regression, was observed as the numbers of codon changes increased. This work suggests that codon usage may modify protein production in ways that influence disease outcome and that evaluation of synonymous codons should be included in the analysis of genetic variants of infectious agents and their association with disease.

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In vivo analysis of mRNA stability using the Tet-Off system in the chicken embryo

Cited by 38 publications

References 60 publications

A region of 35 kb containing the trace amine associate receptor 6 (TAAR6) gene is associated with schizophrenia in the Irish study of high-density schizophrenia families

A region of 35 kb containing the trace amine associate receptor 6 (TAAR6) gene is associated with schizophrenia in the Irish study of high-density schizophrenia families

Rediscovering the chick embryo as a model to study retinal development

Synonymous codon changes in the oncogenes of the cottontail rabbit papillomavirus lead to increased oncogenicity and immunogenicity of the virus

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