Genomic and metabolic engineering + cellular agriculture

Question 1

What genetic disease leads to low T or killer cell levels?

Answer 1

SCIDX1

Question 2

A functioning IL-2RG gene would be delivered to ______ cells via a ______ vector to treat ______.

Answer 2

A functioning IL-2RG gene would be delivered to hematopoietic stem cells via a retrovirus vector to treat SCIDX1.

Question 3

Why was the integration of IL-2RG harmful to some patients?

Answer 3

The retrovirus integrated near an oncogene.

Question 4

What is the advantage of non-viral vectors over viral vectors?

Answer 4

Cheaper and safer.

Question 5

What is the advantage of viral vectors over non-viral vectors?

Answer 5

Higher transfection efficiency.

Question 6

What kind of mutagenesis is risked with viral vectors?

Answer 6

Insertional mutagenesis.

Question 7

What are the two ways in which eukaryotic cells repair DSBs?

Answer 7

1. Homology-directed repair
2. Non-homologous end joining

Question 8

What is the typical ‘donor template’ for homology-directed repair in a eukaryotic cell?

Answer 8

The homologous chromosome.

Question 9

For gene knock-out, would we use HDR or NHEJ?

Answer 9

NHEJ - random mutagenesis.

Question 10

What type of enzyme would introduce highly specific DSBs?

Answer 10

Meganucleases - target larger (12-40bp) recognition sites.

Question 11

Zinc finger is a ___-containing protein that can bind to ___/___.

Answer 11

Zinc finger is a zinc-containing protein that can bind to DNA/RNA.

Question 12

Where are ZF domains found in nature?

Answer 12

Transcription factors.

Question 13

How many bases does one ZF domain bind?

Answer 13

3 bases.

Question 14

What does ZFN stand for? How many bases does a ZFN target?

Answer 14

Zinc-finger nuclease - containing three ZF domains which thus bind a 9 base sequence.

Question 15

What does TALEN stand for?

Answer 15

Transcription activator-like effector nucleases.

Question 16

What DNA motif do TALENs target?

Answer 16

TAL effector motifs.

Question 17

What are three advantages of turning cells into ‘factories’ via metabolic engineering?

Answer 17

1. Cheap and efficient.
2. Highly engineerable.
3. Can perform difficult chemistry.

Question 18

A series of enzymes that converts a substrate to a final product is called a…

Answer 18

Metabolic pathway.

Question 19

What are the two main methods of metabolic engineering?

Answer 19

1. Introducing exogenous enzymes to a pathway.
2. Using recombinant DNA to edit a pathway.

Question 20

Why may we need substrate channeling when engineering a metabolic pathway?

Answer 20

Exogenous enzymes may interfere adversely with other host enzymatic pathways.

Question 21

What is substrate channeling?

Answer 21

Taking the product of one enzyme and transferring it directly to another enzyme.

Question 22

We can physically join enzymes together for substrate channeling with…

Answer 22

An enzyme scaffold.

Question 23

Why would we have additional copies of an enzyme in a scaffold?

Answer 23

If it is a rate-limiting enzyme, more copies prevent a bottleneck in the pathway.

Question 24

Why might nanoparticles be useful in metabolic engineering?

Answer 24

Nanoparticles could compartmentalise enzymatic reactions with toxic intermediates.

Question 25

What are three advantages of cultivated meat?

Answer 25

1. Less contamination.
2. Less antibiotics/hormones/microplastics.
3. Remove animal killing.
4. Food security.

Question 26

What are organoleptics?

Answer 26

Compounds that create the aroma/taste/texture of a food.

Question 27

What are the three technology classes of meat cultivation?

Answer 27

1. Cell line development
2. Media development
3. Scaffolding