Genomic and metabolic engineering + cellular agriculture Flashcards
What genetic disease leads to low T or killer cell levels?
SCIDX1
A functioning IL-2RG gene would be delivered to ______ cells via a ______ vector to treat ______.
A functioning IL-2RG gene would be delivered to hematopoietic stem cells via a retrovirus vector to treat SCIDX1.
Why was the integration of IL-2RG harmful to some patients?
The retrovirus integrated near an oncogene.
What is the advantage of non-viral vectors over viral vectors?
Cheaper and safer.
What is the advantage of viral vectors over non-viral vectors?
Higher transfection efficiency.
What kind of mutagenesis is risked with viral vectors?
Insertional mutagenesis.
What are the two ways in which eukaryotic cells repair DSBs?
- Homology-directed repair
- Non-homologous end joining
What is the typical ‘donor template’ for homology-directed repair in a eukaryotic cell?
The homologous chromosome.
For gene knock-out, would we use HDR or NHEJ?
NHEJ - random mutagenesis.
What type of enzyme would introduce highly specific DSBs?
Meganucleases - target larger (12-40bp) recognition sites.
Zinc finger is a ___-containing protein that can bind to ___/___.
Zinc finger is a zinc-containing protein that can bind to DNA/RNA.
Where are ZF domains found in nature?
Transcription factors.
How many bases does one ZF domain bind?
3 bases.
What does ZFN stand for? How many bases does a ZFN target?
Zinc-finger nuclease - containing three ZF domains which thus bind a 9 base sequence.
What does TALEN stand for?
Transcription activator-like effector nucleases.
What DNA motif do TALENs target?
TAL effector motifs.
What are three advantages of turning cells into ‘factories’ via metabolic engineering?
- Cheap and efficient.
- Highly engineerable.
- Can perform difficult chemistry.
A series of enzymes that converts a substrate to a final product is called a…
Metabolic pathway.
What are the two main methods of metabolic engineering?
- Introducing exogenous enzymes to a pathway.
- Using recombinant DNA to edit a pathway.
Why may we need substrate channeling when engineering a metabolic pathway?
Exogenous enzymes may interfere adversely with other host enzymatic pathways.
What is substrate channeling?
Taking the product of one enzyme and transferring it directly to another enzyme.
We can physically join enzymes together for substrate channeling with…
An enzyme scaffold.
Why would we have additional copies of an enzyme in a scaffold?
If it is a rate-limiting enzyme, more copies prevent a bottleneck in the pathway.
Why might nanoparticles be useful in metabolic engineering?
Nanoparticles could compartmentalise enzymatic reactions with toxic intermediates.
