Gene trapping—a preface

Evans, Martin

doi:10.1002/(sici)1097-0177(199806)212:2<167::aid-aja1>3.3.co;2-5

Cited by 8 publications

(8 citation statements)

References 5 publications

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“…However, as has been found in homologous-recombination experiments, there are 'hot' and 'cold' genomic spots for gene-trap vector insertions. But, computer modelling, albeit with a limited data set, has indicated that on the basis of the percentage of unique clones that have been trapped, and given enough rounds of mutagenesis and enough different types of vector to negate biases, virtually the entire ES-cell genome could be saturated with insertions 36 . This modelling exercise needs to be repeated using the new data sets from the large genotype-driven screens; however, data from the German Gene Trap Consortium, which has used four different vectors to isolate ~6,000 sequences that represent ~4,500 genes (BOX 2) (W. Wurst, personal communication), are consistent with this model.…”

Section: New Vector Designsmentioning

confidence: 99%

Gene-trap mutagenesis: past, present and beyond

2001

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Although at least 35,000 human genes have been sequenced and mapped, adequate expression or functional information is available for only approximately 15% of them. Gene-trap mutagenesis is a technique that randomly generates loss-of-function mutations and reports the expression of many mouse genes. At present, several large-scale, gene-trap screens are being carried out with various new vectors, which aim to generate a public resource of mutagenized embryonic stem (ES) cells. This resource now includes more than 8,000 mutagenized ES-cell lines, which are freely available, making it an appropriate time to evaluate the recent advances in this area of genomic technology and the technical hurdles it has yet to overcome.

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Section: New Vector Designsmentioning

confidence: 99%

Gene-trap mutagenesis: past, present and beyond

2001

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“…In this context, gene trapping in embryonic stem (ES) cells provides a tagged insertional mutagenesis suited for large‐scale studies (reviewed by Zambrowicz and Friedrich, 1998). Gene trapping by random insertional mutagenesis tags genes in ways that allows facile determination of their expression pattern and DNA sequence, as well as the generation of mouse mutants (Gossler et al, 1989; reviewed by Evans et al, 1997; Evans, 1998).…”

Section: Introductionmentioning

confidence: 99%

Sorting nexin‐14, a gene expressed in motoneurons trapped by an in vitro preselection method

Carroll

Renoncourt

Gayet

et al. 2001

Developmental Dynamics

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A gene-trap strategy was set up in embryonic stem (ES) cells with the aim of trapping genes expressed in restricted neuronal lineages. The vector used trap genes irrespective of their activity in undifferentiated totipotent ES cells. Clones were subjected individually to differentiation in a system in which ES cells differentiated into neurons. Two ES clones in which the trapped gene was expressed in ES-derived neurons were studied in detail. The corresponding cDNAs were cloned, sequenced, and analysed by in situ hybridisation on wild-type embryo sections. Both genes are expressed in the nervous system. One gene, YR-23, encodes a large intracellular protein of unknown function. The second clone, YR-14, represents a sorting nexin (SNX14) gene whose expression in vivo coincides with that of LIM-homeodomain Islet-1 in several tissues. Sorting nexins are proteins associated with the endoplasmic reticulum (ER) and may play a role in receptor trafficking. Gene trapping followed by screening based on in vitro preselection of differentiated ES recombinant clones, therefore, has the potential to identify integration events in subsets of genes before generation of mouse mutants.

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“…Gene trapping, on the other hand, is a form of random insertional mutagenesis specifically designed to disrupt and retrieve genes in target cells by producing intragenic vector integration events (4,5). Hundreds or even thousands of genome sequences in mouse ES cells can be ''trapped'' in a relatively short period (6,7).…”

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confidence: 99%

Expression profiling with arrays of randomly disrupted genes in mouse embryonic stem cells leads to in vivo functional analysis

Matsuda

Shigeoka²,

Iida³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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DNA arrays are capable of profiling the expression patterns of many genes in a single experiment. After finding a gene of interest in a DNA array, however, labor-intensive gene-targeting experiments sometimes must be performed for the in vivo analysis of the gene function. With random gene trapping, on the other hand, it is relatively easy to disrupt and retrieve hundreds of genes͞gene candidates in mouse embryonic stem (ES) cells, but one could overlook potentially important gene-disruption events if only the nucleotide sequences and not the expression patterns of the trapped DNA segments are analyzed. To combine the benefits of the above two experimental systems, we first created Ϸ900 genetrapped mouse ES cell clones and then constructed arrays of cDNAs derived from the disrupted genes. By using these arrays, we identified a novel gene predominantly expressed in the mouse brain, and the corresponding ES cell clone was used to produced mice homozygous for the disrupted allele of the gene. Detailed analysis of the knockout mice revealed that the gene trap vector completely abolished gene expression downstream of its integration site. Therefore, identification of a gene or novel gene candidate with an interesting expression pattern by using this type of DNA array immediately allows the production of knockout mice from an ES cell clone with a disrupted allele of the sequence of interest.

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Gene trapping—a preface

Abstract: Fig. 1. The average number of unique clones found per 1,000 picked for a variety of total genome target sizes and for between 0% and 20% of the loci being hit at 1-20 times normal.

Cited by 8 publications

References 5 publications

Gene-trap mutagenesis: past, present and beyond

Gene-trap mutagenesis: past, present and beyond

Sorting nexin‐14, a gene expressed in motoneurons trapped by an in vitro preselection method

Expression profiling with arrays of randomly disrupted genes in mouse embryonic stem cells leads to in vivo functional analysis

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