#151632

RalGDS KO Mouse

Cat. #151632

RalGDS KO Mouse

Cat. #: 151632

Sub-type: Mouse

Availability: 6-8 weeks

Disease: Melanoma

Model: Knock-Out

This fee is applicable only for non-profit organisations. If you are a for-profit organisation or a researcher working on commercially-sponsored academic research, you will need to contact our licensing team for a commercial use license.

Contributor

Inventor: Chris Marshall

Institute: The Institute of Cancer Research

Tool Details
Handling
Target Details
Related Tools
References

Tool Details

*FOR RESEARCH USE ONLY

  • Tool name: RalGDS KO Mouse
  • Research fields: Cancer;Cell signaling and signal transduction;Genetics
  • Tool sub type: Mouse
  • Disease: Melanoma
  • Model: Knock-Out
  • Conditional: No
  • 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).
  • Genetic background: 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
  • 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.

Handling

  • Shipping conditions: Embryo/Spermatoza- Dry Ice

Target Details

  • Target: RalGDS

Related Tools

  • Related tools: RalGDS Floxed Mouse

References

  • Gonzlez-Garca et al. 2005. Cancer Cell. 7(3):219-26. PMID: 15766660.
  • RalGDS is required for tumor formation in a model of skin carcinogenesis.