mouse models

Question 1

why use mice?

Answer 1

• Small domesticated animal
• Same organ and tissue systems than humans
• Generation time short: 3 months
• Large litter size: 7-10 pups
• Germline can be modified via genetic engineering

Question 2

the 3R principle

Answer 2

1. replace (with computer or cell model)
2. reduce (smallest number of animals)
3. refine (optimization of animal treatment)

Question 3

types of effects

Answer 3

• Loss of expression
• Increase expression
• Change sequence of target gene

Question 4

how to implement the effects

Answer 4

1. vector with DNA –> homologous recombination
2. Crispr-Cas9

Question 5

modification types

Answer 5

• stable
• conditional
• ineducable

Question 6

pipeline - mouse model for McArdle disease

Answer 6

1. vector design
2. electroporation of stem cells
3. long PCR to identify recombinant clones
4. southern blot to identify homologous recombination
5. karytoping to exclude fusings etc.
6. mouse blastocyst injection
7. corssing with CRE strain