Synthetic Biology Flashcards
What are the two beliefs for the meaning of synthetic life?
- Synthetic cell is one that operates off a chemically synthesised genome.
- In vitro life as where biochemical reactions placed together can carry out the function of life and act as artifical cells
Why do we want to make synthetic cells?
More predictable than natural cells that we don’t fully understand.
Can be streamlined for one task only (use fewer resources).
Easily confine the cells (conditional growth only in lab environment)
How was the first synthetic cell (Synthia) developed?
Assemble overlapping synthetic DNA olignucleotides into cassettes - assembled by homologous recombination into completely assembled synthetic ds genome.
Genome injected into recipient cell (genome transplantion) and synthetic cell outgrows natural host.
Hardware is difficult to synthetically manufacture so chemically synthesise gene and transplant into donor Mycoplasma capricolum cell which rebooted it as M. mycoides capable of self-replication
What is the difference between parts, devices and systems?
Parts encode biological functions - promoter, ribosome binding sites, protein coding sequences, terminators.
Devices made from collection of parts and encode human-defined functions - protein-generator, inverter, receiver, synthetic genetic toggle switches.
Systems are one or more devices designed to perform tasks - e.g. synthesiser, sensor, seeker and striker systems
Describe DNA synthesis as a tool in synthetic biology.
Agilent technologies oligo synthesis via inkjet printing. Multiple layers of nucleotides precisely printed and added to chain. Getting cheaper, easier and faster
Describe DNA sequencing as a tool in synthetic biology.
Sanger Sequencing
Next gen sequencing are high throughput (much faster, cheaper) - most, except nanopore, are sequencing by synthesis approaches and involve solid-phase amplification of individual clonal templates.
Bench-top sequencing platforms generate data quickly and cheaply.
Describe DNA assembling as a tool in synthetic biology.
Traditional way is by finding restriction sites and cloning into DNA.
Gibson assembly using overlapping sequences
Add to pot with T5 exonuclease, Taq Ligase and Phusion polymerase for 30 minutes at 50dc.
Exonuclease chews back 5’ ends, fragments anneal, DNA polymerases closes gaps, DNA ligase seals nicks
What are the applications of synthetic biology?
Microbial cell factories - protein products, insulin, fuels, fine/specialty chemicals and drugs.
Medical and health - biosensor, smart therapies, treatment of metabolic disorders, tissue engineering
Sustainable world - bioremediation, biomining, engineering crops and commensal soil organism
