Functional Genome

Question 1

What is whole genome sequencing for?

Answer 1

→ Used to capture the sequence of the coding region of the genome

Question 2

How do you prove that a gene variant causes dysfunction?

Answer 2

→ Knock out gene

→ over expression of gene

Question 3

How can primary cells be kept?

Answer 3

→ they can be immortalised

Question 4

How do you filter out mutations in WES?

Answer 4

→ remove the synonymous mutations
→filter out variants in databases that are common
→look at family members genotypes

Question 5

What are the assumptions in WES?

Answer 5

→ The variant is within the coding region

Question 6

Why is only isolating a gene not sufficient for a diagnosis?

Answer 6

→ you need to prove that the gene causes disease

Question 7

Why are biopsies of affected tissues not always available?

Answer 7

→ Gene isnt expressed in the blood

→ tissue is not accessible

Question 8

What does in vitro mean?

Answer 8

→ Removal of cells from an animal and subsequent growth in favorable conditions

Question 9

What is an advantage of in vitro?

Answer 9

→ Cheap
→ rapid
→ reproducible model

Question 10

Describe how RNAi gene knockdown works?

Answer 10

→ a sequence is introduced that is complementary to the gene of interest
→ it is packaged within a plasmid and contains a promoter (RNA polymerase III)
→ they are transfected into the nucleus
→ the RNA is transcribed and exits the nucleus through the pores with exportin 5 protein
→ It then is cleaved by Dicer
→ it leaves the complementary DNA
→ the DNA binds to the RISC complex
→ and this seeks the gene of interest RNA and silences it by cutting it

Question 11

WHat is the effect of knockdown on PDZRN3?

Answer 11

→ Inhibition of myotube formation and MHC expression