Plasmids for Optogenetics Research
The field of optogenetics integrates optics and genetic engineering approaches and technology to measure and manipulate cells and their governing biomolecular processes. The tools and technologies developed for optogenetics research utilize light to detect, measure, and control molecular signals, cells, and groups of cells in order to understand their activity and the effects of alterations to this activity.
These genetically-encoded tools have come to represent two functions in optogenetics: actuators (genetically-encoded tools for light-activated control of neurons; e.g., microbial opsins) and sensors (genetically-encoded reporters of molecular signals; e.g., calcium indicators). We have organized the optogenetics plasmids available in our repository into actuators and sensors below.
Interested in more news about optogenetics research? Check out the related posts on Addgene's blog.
Plasmids for Optogenetics Actuators
Addgene's repository contains a variety of microbial opsins and additional tools to control neuronal activity. These optogenetics actuators have been organized based on the opsin(s) or tool encoded by the plasmid. Click on the link to jump to plasmids which encode Channelrhodopsins, Halorhodopsins, Archaerhodopsins, Leptosphaeria rhodopsins, or other actuator tools for protein localization, controlled gene expression, and more.
|ChR2||Light-gated cation channel (from Chlamydomonas reinhardtii )||470||GFP|
|ChR2 mutants||ChETA (ChR2 with mutations at position 123)||EYFP|
|ChR2 with various mutations||EYFP|
|VChR1||Light-gated cation channel (from Volvox carteri )||540-570||EYFP|
|ChIEF||ChR1-ChR2 chimera with site-directed mutagenesis||470||tdTomato|
|V1C1||ChR1-VChR1 chimera with E122T and E162T mutations||540||EYFP|
|ChR2-2A-Halo||Chr2-GFP and Halo-YFP fused with self-cleaving 2A linker||470/589||GFP/YFP|
SlowChloC: Chloride-conducting cation, ChR2 modification: mutations E90R, D156N, T159C, slow kinetics
ChloC: Chloride-conducting cation, ChR2 modification: mutations E90R, T159C, fast kinetics
|iChloC||Improved ChloC; light-gated chloride channel; AAV backbone; slowChloC mutations plus 2 additional amino acid substitutions (E83Q & E101S)||465||tdimer2|
|SwiChRca||Chloride-conducting cation, iC1C2 modification: mutations C128A||475 (activ.) / 632 (inactiv.)||EYFP|
|Chronos||Blue- and green-light drivable (from Stigeoclonium helveticum )||470, 530 (activ.)||GFP|
|Chrimson||Red-light drivable (from Chlamydomonas noctigama )||590 (activ.)||GFP|
|ChrimsonR||K176R mutant of Chrimson||590 (activ.)||tdTomato|
|GtACR||Guillardia theta anion channel rhodopsins|
|Jaws||Red-shifted, light-driven inward chloride pump, used to silence neuronal activity (from Haloarcula (Halobacterium) salinarum (strain Shark); Cruxhalorhodopsin class||632||GFP|
|Halo (aka NpHR)||Light-driven inward chloride pump, used to silence neuronal activity (from Natronomonas pharaonis )||589||GFP|
|eNpHR (aka NpHR)||Light-driven inward chloride pump, used to silence neuronal activity (from Natronomonas pharaonis )||589||EYFP|
|eNpHR 3.0||eNpHR with addition of trafficking signal from Kir2.1 and ER export signal||589||EYFP|
|Arch||Light-activated outward proton pump (from Halorubrum sodomense )||566||GFP|
|eArch 3.0||Arch with addition of trafficking signal from Kir2.1||566||EYFP|
|ArchT||Light-activated outward proton pump (from Halorubrum genus )||566||GFP|
|eArchT 3.0||ArchT with addition of trafficking signal from Kir2.1||566||EYFP|
|Mac||Light-activated outward proton pump (from Leptosphaeria maculans )||540||GFP|
|eMac 3.0||Mac with addition of trafficking signal from Kir2.1||540||EYFP|
Protein Localization, Controlled Gene Expression, and More
|bPAC||Light-activated adenylyl cyclase (from Beggiatoa sp.) for light-induced cAMP modulation|
|BphP1–PpsR2||Near IR induced binding can be used to translocate target proteins to specific cellular compartments or change gene expression.||Venus, mCherry|
|Cryptochrome 2 and CIB1||Blue-light–mediated induction of protein interactions based on Arabidopsis thaliana cryptochrome 2 and CIB1||EGFP, mCherry|
|Dronpa||Light-inducible Dronpa mutant domains that associate and cage a protein in the dark, while dissociate and activate the protein in the light||~400nm (to switch on); ~500nm (to switch off)|
|iLID||Improved light-inducible dimers; Bacterial SsrA peptide is embedded in the LOV2 domain (iLID plasmid), while its natural binding partner SspB is included in a separate plasmid; Blue light activation induces a conformational change in the LOV2 domain which allows SsrA to bind with SspB||450||
|LAD||Light-activated dimerization, using proteins GIGANTEA (GI) and the light oxygen voltage (LOV) domain of FKF1||450|
|LITEZ||Light-inducible transcription using engineered zinc finger proteins, using proteins GIGANTEA (GI) and the light oxygen voltage (LOV) domain of FKF1||450|
|Lumitoxin||Peptide toxin for blocking K + (Kv) channels fused to LOV2 photoswitch||mCherry|
|pDawn||Blue-light photoreceptor for light-activated gene expression||470|
|pDusk||Blue-light photoreceptor for light-repressed gene expression||470|
|Phy–PIF||Light-controlled PhyB–PIF6 interaction for reversible translocation of proteins||mCherry, mYFP|
|Rac1-LOV||Rac1 fused to photoreactive LOV for localized, reversible Rac activation to control cell motility||mCherry, mVenus|
|TULIPs||Tunable light-inducible dimerization tags (TULIPs), plasmids allow cloning of protein coding sequences with GFP-LOVpep, cpPDZ, ePDZb and ePDZb1 as tags||458- or 473-nm||GFP, mCherry|
Plasmids for Optogenetics Sensors
Addgene's repository contains a variety of genetically encoded biosensors to monitor and measure fluctuations in molecular signals in cells, including those to monitor calcium, glutamate, pH, and voltage. These optogenetics sensors have been organized below into plasmids containing calcium sensors and other sensors .
|GCaMP||GCaMP2||Modified calcium indicator derived from a fusion of a cpEGFP with C-terminus CaM and N-terminus M13 (GCaMP)|
|GECO||G-GECO1.0||Genetically encoded calcium indicator (GECO)|
|Twitch||Twitch-1||High-performing calcium indicator based on C-terminal domain of troponin C|
|Chloride & pH||ClopHensor||Simultaneous, measurement of Cl - and pH with E 2 GFP indicator|
|ClopHensorN||Simultaneous measurement of Cl- and pH optimized for use in the nervous system|
|Glutamate||iGluSnFR||Intensity-based glutamate-sensing fluorescent reporter (iGluSnFR)|
|pH||sypHy||Fluorescent reporter of pH-sensitive pHluorin fused to synaptophysin|
|Voltage||VSFP||VSFP2.1||Voltage-sensitive fluorescent protein (VSFP); Ci-VSP voltage sensor domain fused to a pair of fluorescent proteins (C-terminally truncated CFP and YFP)|
|VSFP3.1||Modified VSFP with R217Q mutation in the S4 domain of Ci-VSP|
|VSFP-CR||VSFP fused to Clover-mRuby2 FRET pair|
|ArcLight||ArcLight-S249||Modified ArcLight (voltage-sensing domain of Ci-VSP and super ecliptic pHluorin with A227D mutation) with pHluorin at position S249 in Ci-VSP|
|ArcLight-Q239||Modified ArcLight with pHluorin at position Q239 in Ci-VSP|
|ArcLight-A242||Modified ArcLight with pHluorin at position A242 in Ci-VSP|
|ASAP||ASAP1||Accelerated Sensor of Action Potentials 1 (ASAP1), a cpGFP is inserted in an extracellular loop of a voltage-sensing domain|
Boyden, et al, Nat Neurosci 2005 Sep; 8(9):1263-8.
Deisseroth et al., J Neurosci 2006 Oct 11; 26(41):10380-6.
Dugué GP et al., Prog Brain Res 2012; 196: 1-28.
Alford SC et al., Biol Cell 2013 Jan; 105(1): 14-29.
OpenOptogenetics: the optogenetics wiki.
Edward Boyden's Lab: Optogenetics Resources
Karl Deisseroth's Lab:< a href="http://www.stanford.edu/group/dlab/optogenetics/" target="blank">Optogenetics Resource Center