F9 Strain improvement by metabolic engineering

Question 1

Genetic (metabolic) engineering include:

Answer 1

▪ Redirection of metabolic pathways towards products of interest
▪ Expanding substrate specificities and increasing robustness

Question 2

design

Answer 2

pathway selection

turn genes on/off

choosing host organisms

expression system

Question 3

Pathway assembly and strain construction (yeast)

Answer 3

• Genetic parts 
- Synthesized, PCR amplified 
• Parts assembly (cloning)
- In bacteria (shuttle vectors)
- In yeast (in vivo recombination) 
• Integration (in vivo recombination) or plasmid

Question 4

CRISPR/Cas9 based genome editing

Answer 4

1. The Cas9 protein: ”The scissors” - cutting
2. The sgRNA: “The address” – where to cut
3. The dDNA: “The bandage” – repairing
   - If no dDNA, the cut can be repaired by NHEJ