Optogenetics

Question 1

Describe optogenetics

Answer 1

• Implant into nerve cells photosynthetic molecules

- Now you can turn light into electricity, so you shine lasers to turn neurons on and off

Question 2

Why can optogenetics not be used in humans?

Answer 2

• Requires a gene therapy to deliver the gene into the nerve

- Molecules come from algae and bacteria, and so they can be attacked by the immune system

Question 3

Describe AAV transfer plasmids

Answer 3

• Transfer plasmid contains AAV, encoding GFP.
• ITRs identify AAV plasmids, as they have 2 ITRs. ITRs and what is in between is included into the vector
• Transgene is the gene you are study and want to get expressed.
• Promotors are specific to organisms and sometimes even tissue types. They can also induce different levels of expression
• Lox sites are 34 base sequences, recognised by regongrinase Cre. These turn genes on and off.
• Cis regulatory elements affect how well a gene is expressed (eg. poly A and WPRE, just upstream of the final ItR)
• PolyA prevents degredation of RNA, while WRPE increases nuclear export

Question 4

Describe actuators

Answer 4

• When retinol binds to opsin the retinal opsin complex becomes activated
• Channelrhodopsin-2
• Chloride pumps (halorhodopsin, chloride pumps therefore they are inhibitory)