Synthetic Biology

Question 1

What is a minimal cell?

Answer 1

A cell that has the minimal genetic functions essential for life.

Question 2

What was the first synthetic cell?

Answer 2

Synthia 1.0 - it took 15 years and $50 million to make it

It was completed in 2010.

Question 3

What was the first synthetic minimal cell?

Answer 3

Synthia 3.0. It was done by minimising Synthia 1.0 using global transposon mutagenesis to identify the essential genes.
It was whittled down to 473 genes.

Question 4

What is a part and what is a device?

Answer 4

Part - something that encodes biological function. Eg. Promoter.

Device - a collection of parts that encode functions. Eg. Genetic toggle switches.

Question 5

Give 4 engineering tools

Answer 5

DNA synthesis
DNA sequencing
DNA assembly
Genome editing/engineering

Question 6

How do you do DNA synthesis?

Answer 6

It is done in 4 steps:
1- Deprotection (DMP protection molecule is removed by acid catalysed enzymes so that a base can be added)
2- Base coupling (a base is added using a tetrazole activator).
3- Capping (this is optional) (unreacted -OH are acetylated to prevent further chain extension).
4- Oxidation (oxydation of phosphate using iodine).

This can be done via Agilent Microarray Technology (inkjet printing).

Question 7

How do you do DNA sequencing?

Answer 7

Sanger sequencing
Or
Next generation sequencing (454 Roche, Illumina, Ion torrent, pac Bio).

Question 8

How do you do DNA assembly?

Answer 8

You put the fragments of DNA together using BioBrick-2 Standard Assembly or Gibson isothermal assembly.

Question 9

How do you do Genome Editing/Engineering?

Answer 9

Can use the CRISPR/Cas9 system or Multiplex automated genomic engineering (MAGE).