Flashcards in Applications of genetic engineering - lecture 13 Deck (12)
What are the differences between stable and transient transformations?
Transient transfections - short term examinations
stable transfections -allow larger term investigations
How is a transient transfection of a celle done?
- using a lysosomes , with genetic material o the inside
- can use reporter genes to deterine if the genetic material has been taken up such as GFP
What is the significant difference bewtween stable and transient transfections?
stable - DNA incorporated in to the genome
transient - DNA not incorporated into the genome
How can stable transfection occur?
By retrovirus infection
How can you make a retro virus ?
Via a packaging cell- expresses all viral proteins and RNA for the virus to be produced
What is significant about a retrovirus?
Contains reverse transcriptase , so the RNA can be turned into to DNA and incorporated in to the genome
How can you get rid of single genetic mutations?
- Take a slection marker in a new piece of chromosome
- allow the chromosome with the targeted gene and selection marker to go through homologous recomibnation
- selection marker has replaced the mutated sequence
How do you make a chimeric mouse?
- embryonic stem cells from balck mouse , induce modified gene
- These genetically mutated ' black mouse cells' are injected into the blastocyst stage of the embryo from a brown mouse
- Impant embryo into surrogate mother , mother gives birth to a chimeric pup
How can you delete a gene in a specific tissue?
Use the cre- Loxp methodology
How is Haemophilia A caused?
mutations in the gene encoding factor VIII , which is located ono X chromosome
What does Hameophilia A cause?
Allows bleeding to continue as blot clots cannot form