Gene trapping and functional genomics

Evans, Martin; Carlton, Mark; Russ, Andreas

doi:10.1016/s0168-9525(97)81166-3

Cited by 20 publications

(22 citation statements)

References 21 publications

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“…Thus, the expression of the reporter gene mimics the expression of the endogenous gene, and the insertion of the vector has the potential to mutate the locus (Evans et al 1997). The use of embryonic stem (ES) cells can generate mutant mice far more quickly than conventional homologous recombination because there is no need to prepare each construct and to target each gene separately.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Ayu17-449 gene expression and resultant kidney pathology in a knockout mouse model

Tang

Araki

et al. 2008

Transgenic Res

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The gene trap technique is a powerful approach for characterizing and mutating genes in the mouse. We used this method to identify a mouse gene of unknown function and to establish a mutant mouse line. We subsequently identified one gene, denoted Ayu17-449, on mouse chromosome 3 that comprised 14 exons encoding 1920 amino acids with a granin motif in its N-terminal sequence. In adult mice, this gene was highly expressed in the brain, heart, lung, muscle, stomach, and kidney. The insertion of a trap vector into the second intron of this gene resulted in the null mutation. Homozygous mice for these mutation died by 1 day after birth. Mutant mice showed a loss of acidic granules in the proximal convoluted tubules of the kidney. Our data demonstrates that Ayu17-449 is important for mouse survival.

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Section: Introductionmentioning

confidence: 99%

Characterization of Ayu17-449 gene expression and resultant kidney pathology in a knockout mouse model

Tang

Araki

et al. 2008

Transgenic Res

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“…In mice, IRESs have been used in "gene-trap" vectors (Skarnes et al, 1992;Evans et al, 1997) to increase the efficiency of gene trapping (Mountford and Smith, 1995;Chowdhury et al, 1997) by eliminating the requirement for in-frame fusion to the interrupted locus. In preliminary experiments using the Sleeping Beauty transposon system (Ivics et al, 1997), the EMCV IRES along with a GFP gene can act as a powerful gene trap in zebrafish (Clark, K.J., et al, unpub.…”

Section: Discussionmentioning

confidence: 99%

Dicistronic Gene Expression in Developing Zebrafish

et al. 1999

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Internal ribosome entry sites (IRESs) allow ribosomal access to messenger RNA without a requirement for cap recognition and subsequent scanning to an initiator AUG. Hence, IRESs have been adapted into dicistronic vectors for the expression of more than one gene from a single mRNA. Dicistronic vectors have been used for many applications in mammalian tissue culture and transgenesis. However, whether the IRESs from mammalian viruses function without temporal or spatial restrictions in nonmammalian organisms like zebra fish (Danio rerio) is unknown. Therefore, we have examined the expression capabilities of the encephalomyocarditis virus (EMCV) IRES during zebrafish embryogenesis. We determined that the EMCV IRES was sufficient to permit detectable expression of several second cistron reporters during zebrafish embryogenesis, including luciferase and green fluorescent protein. This suggests that our dicistronic vectors are suitable for general use in any vertebrate system, from fish to humans.

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“…The IMMC proposes combining both genotype and phenotype-driven approaches to achieve this goal. The genotype-driven approach employs ES-cell technology to create targeted mutations by gene knock-out, knock-in or gene-trapping (Evans et al, 1997;Roths et al, 1999). This method has helped define the biological activities of many genes.…”

Section: Introductionmentioning

confidence: 99%