EXPERIMENTAL MODELS
Contributor Information
- Name Chris Marshall
- Institute The Institute of Cancer Research
Tool Details
- Tool name: RalGDS KO Mouse
- Tool type: Experimental models
- Tool sub-type: Mouse
- Disease: Melanoma; GEF; Ral; RalGDS; Ras
- Model: Knock-Out
- Conditional: No
- Genetic background and cross history: Mouse RalGDS genomic clones were obtained by screening a 129/SvJ BAC library (Incyte). A targeting vector was designed using a DNA fragment extending from intron 7 to the 3'UTR, cloned into the pKO scrambler 901 vector (Stratagene). A neomycin cassette flanked by two loxP sites was cloned upstream of exon 16 and a 3rd loxP site was located in exon 8. The exons flanked by loxP sites comprise part of the catalytic domain of RalGDS and residues involved in the binding of the exchange factor to Ras. The construct was electroporated into RW4 ES cells (Incyte) and clones positive for recombination were then transiently transfected with pcrePac vector to eliminate the neomycin cassette. Cells carrying the RalGDS allele lacking exons 9-15 were injected into MF-1 blastocysts and germline-transmitting chimeric mice were obtained. Animals with a mixed MF-1/129SvJ background were used throughout. Intercrossing of RalGDS-/+ mice yielded the expected Mendelian ratios, indicating that no embryonic lethality had occurred. Moreover, RalGDS-/- male and female mice are fertile and no major defects in any organs studied have been observed.
- Phenotype: RalGDS-/- mice show normal development and fertility but reduced tumour development in an established model of chemically-induced skin carcinogenesis.
- Zygosity: Homozygous
- Description: The knock-out mouse was developed by Chris Marshall at the ICR and used to demonstrate a significant role for RalGDS in Ras-dependent carcinogenesis in vivo. Lack of RalGDS resulted in reduced tumour incidence, size and progression to malignancy in multistage skin carcinogenesis, and reduced transformation by Ras in tissue culture. Experiments performed in cells isolated from skin tumours suggested that RalGDS mediates cell survival through the activation of the JNK/SAPK pathway. Mouse deficient in RalGDS (a member of the RalGEF family, which control the activity of the small GTPases RalA and RalB, and also have Ras binding domains).
- Research area: Cancer; Cell signaling and signal transduction; Genetics
- Production details: Mouse RalGDS genomic clones were obtained by screening a 129/SvJ BAC library (Incyte). A targeting vector was designed using a DNA fragment extending from intron 7 to the 3'UTR, cloned into the pKO scrambler 901 vector (Stratagene). A neomycin cassette flanked by two loxP sites was cloned upstream of exon 16 and a 3rd loxP site was located in exon 8. The exons flanked by loxP sites comprise part of the catalytic domain of RalGDS and residues involved in the binding of the exchange factor to Ras. The construct was electroporated into RW4 ES cells (Incyte) and clones positive for recombination were then transiently transfected with pcrePac vector to eliminate the neomycin cassette. Cells carrying the RalGDS allele lacking exons 9-15 were injected into MF-1 blastocysts and germline-transmitting chimeric mice were obtained. Animals with a mixed MF-1/129SvJ background were used throughout. Intercrossing of RalGDS-/+ mice yielded the expected Mendelian ratios, indicating that no embryonic lethality had occurred. Moreover, RalGDS-/- male and female mice are fertile and no major defects in any organs studied have been observed.
- For Research Use Only
Target Details
- Target: RalGDS
Application Details
Handling
- Shipping conditions: Embryo/Spermatoza- Dry Ice
Documentation
Related Tools
References
- • Gonz
lez-Garc a et al. 2005. Cancer Cell. 7(3):219-26. PMID: 15766660. - • RalGDS is required for tumor formation in a model of skin carcinogenesis.
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