OPTOGENETICS

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  • Created by: molg
  • Created on: 19-05-18 17:45
Reporters
1. Ca2+ indicators, pH indicators, genetically encoded indicators for cAMP, indicators of enzyme activity, indicators of transcriptional events ect
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Actuators
light sensitive ion channels, light sensitive ion pumps, light sensitive G-protein couple receptors, light sensitive enzymes, light-sensitive transcriptional switches
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Optogenetics
combination of optics and genetics- genetically engineer new proteins then express them in relevant cells using cell-specific promoters
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ChR2- channel rhodopsin 2
Light-sensitive ion channels
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all-trans-retinal
a co-factor present in all mammalian cellular preparations- absorbs photons
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CHIEF
variant of ChR2- highly sensitive and fast
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STEP-OPSIN
variant of ChR2- slower kinetics- want to shift excitability of a circuit to a higher/lower state- reduces use of light
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Inhibitory actuators
NpHR- orange light- Cl transport into cell, Arch- yellow light- H+ transport out of the cell
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OPTO alpha-1
light-stimulated GPCR- IP3 pathway
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OPTO beta-2
light-stimulated GPCR- cAMP pathway
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bPAC
light stimulated enzyme- dimerises on light stimulation- generates cAMP
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Blue light- caution
can produce artefacts in vivo- absorbed very quickly- doesn't dissipate very quickly- produces too much heat
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red-shifted optogenetic actuaters
red light spreads better- eg. can pass through the skull to evoke muscle twitches
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VChR1
red-shifted compared to ChR2 (absorption max 535 nm)- can be co-expressed ChR2
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ReaChR
synthetic- combining parts of ChiEF, VChR1 and VChR2- high expressing, red-shifted channel0 can be excited by near infra-red light ( absorption max >600nm)
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Card 2

Front

light sensitive ion channels, light sensitive ion pumps, light sensitive G-protein couple receptors, light sensitive enzymes, light-sensitive transcriptional switches

Back

Actuators

Card 3

Front

combination of optics and genetics- genetically engineer new proteins then express them in relevant cells using cell-specific promoters

Back

Preview of the back of card 3

Card 4

Front

Light-sensitive ion channels

Back

Preview of the back of card 4

Card 5

Front

a co-factor present in all mammalian cellular preparations- absorbs photons

Back

Preview of the back of card 5
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