Foxp3-RFP+AKT3-/- transgenic mouse strain to study the role of AKT3 in stem cells and tumourigenicity
1. Study purpose
Epithelial-mesenchymal transition (EMT) is a key developmental program often activated during cancer invasion and metastasis. AXL is a tyrosine kinase transmembrane receptor that have shown to play a role in EMT. AKT3 is a downstream effector of AXL involved in the maintenance of EMT and stem cell-like activity. This project aims to elaborate the role of AKT3 during EMT in physiological development as well as in tumor development by breeding AKT3 knockout mice. The AKT3 knockout strain have a C57bl6/J background. Therefore, the C57bl6/J strain will be used as control with functional AKT3 expression.
2. Expected harm to animals
The project consist of breeding and acute experiments only where expected pain experienced by the mice is minimal. However, animals will be observed daily, and animals with evidence of distress will be euthanized. This project is an extension of ongoing project (FOTS 9596), hence the breeding schedule is already established. The AKT3 null mice will breed together and unwanted genotypes are therefore not expected.
3. Expected benefits to scientific community and society
AKT3 seems to play a role in tumor development and metastasis. We therefore hypothesize that AKT3 null mice will develop less aggressive tumors and metastases. Elucidating the role AKT3 play in normal physiological development and in cancer will contribute to knowledge and basic understanding for the scientific community as well as potentially providing a role in future cancer treatment for the society at large.
4. Amount of animals and species/strain
Based on our need for the study we anticipate using 1000 AKT3 null animals, and 300 C57bl6/J.
5. Complying with replacement, reduction, and refinement
To understand importance of AKT3 in developing animals it is necessary to perform in vivo studies, as in vitro studies cannot mimic the whole body complexity. The number of animals are reduced by in vitro studies prior to in vivo experiment. Furthermore, animals will be used for both in vivo and ex vivo studies. For breeding, we will use heterozygotes to benefit from increased production of offspring during the experiment. As the colony is established, the breeding will be performed according to the planed experiment in order to reduce the number of animals. The normal state in these animals does not cause pain and no painful procedures will be performed during breeding. In order to select animals with targeted genotype, small biopsies are taken from the ear. This approach allow reduction of animals used in experiments. Homozygous wild type individuals are excluded from the colony, in order to exclude unwanted phenotype and decrease number of not-targeted allele amongst newborn pups. As the colony is established, backcrossing will be performed using C57bl6/J from The Jackson Laboratory with particular focus on limiting inbred mice in order to maintain healthy animals in the colony.
Epithelial-mesenchymal transition (EMT) is a key developmental program often activated during cancer invasion and metastasis. AXL is a tyrosine kinase transmembrane receptor that have shown to play a role in EMT. AKT3 is a downstream effector of AXL involved in the maintenance of EMT and stem cell-like activity. This project aims to elaborate the role of AKT3 during EMT in physiological development as well as in tumor development by breeding AKT3 knockout mice. The AKT3 knockout strain have a C57bl6/J background. Therefore, the C57bl6/J strain will be used as control with functional AKT3 expression.
2. Expected harm to animals
The project consist of breeding and acute experiments only where expected pain experienced by the mice is minimal. However, animals will be observed daily, and animals with evidence of distress will be euthanized. This project is an extension of ongoing project (FOTS 9596), hence the breeding schedule is already established. The AKT3 null mice will breed together and unwanted genotypes are therefore not expected.
3. Expected benefits to scientific community and society
AKT3 seems to play a role in tumor development and metastasis. We therefore hypothesize that AKT3 null mice will develop less aggressive tumors and metastases. Elucidating the role AKT3 play in normal physiological development and in cancer will contribute to knowledge and basic understanding for the scientific community as well as potentially providing a role in future cancer treatment for the society at large.
4. Amount of animals and species/strain
Based on our need for the study we anticipate using 1000 AKT3 null animals, and 300 C57bl6/J.
5. Complying with replacement, reduction, and refinement
To understand importance of AKT3 in developing animals it is necessary to perform in vivo studies, as in vitro studies cannot mimic the whole body complexity. The number of animals are reduced by in vitro studies prior to in vivo experiment. Furthermore, animals will be used for both in vivo and ex vivo studies. For breeding, we will use heterozygotes to benefit from increased production of offspring during the experiment. As the colony is established, the breeding will be performed according to the planed experiment in order to reduce the number of animals. The normal state in these animals does not cause pain and no painful procedures will be performed during breeding. In order to select animals with targeted genotype, small biopsies are taken from the ear. This approach allow reduction of animals used in experiments. Homozygous wild type individuals are excluded from the colony, in order to exclude unwanted phenotype and decrease number of not-targeted allele amongst newborn pups. As the colony is established, backcrossing will be performed using C57bl6/J from The Jackson Laboratory with particular focus on limiting inbred mice in order to maintain healthy animals in the colony.