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Optogenetics, a recently developed technology has been commonly used in the field of neuroscience. It is a neuromodulation method that uses a combination of techniques from optics and genetics to control and monitor the activities of individual neurons in living tissue, even in a freely moving animal. This technology allows us to achieve an unprecedented understanding of neural circuit function, such as neural circuit dysfunction underlying mood disorders, addiction, Parkinson's disease, and the neural circuits mediating normal behavior.
In this technology, neurons are first genetically engineered to express a light-sensitive protein (opsin), which is typically an ion channel, pump, or G protein-coupled receptor. When these engineered neurons are then illuminated with light of the correct frequency they will be transiently activated or inhibited or their signaling pathways will be modulated, depending on the particular kind of opsin that was chosen for expression.
Creative Biogene can offer you various optogenetic tool vectors that can express different opsins to meet your specific needs. Besides, Creative Biogene also can provide you with a related service, AAV packaging service, to accelerate the speed of achieving your research goals.
Highlights of Our Optogenetic Tools
Optogenetics integrates optics and genetic engineering to measure and manipulate cells (typically neurons) and control the biological processes they mediate. The tools and techniques developed for optogenetic research utilize light to detect, measure, and control molecular signals and cells to understand their function.
You can utilize Creative Biogene's various optogenetic tools to design experiments such as:
Case Study 1
Calcium ions (Ca2+), as omnipresent secondary messengers, fulfill critical regulatory roles in a diverse array of cerebral functions, encompassing cognition, emotion, locomotion, and learning and memory. To directly interrogate causal associations between Ca2+ signaling and cellular functions in spatial and temporal dimensions, researchers have hitherto engineered a series of molecular optogenetic technologies intended to manipulate intracellular Ca2+ levels through photostimulation.
Figure 1. The primordial iteration of monSTIM1 was comprised of three constituents: green fluorescent protein (GFP), CRY2(E281A, A9) (hereinafter CRY2), and the cytosolic STIM1 fragment. Substituting GFP with a small tag significantly decreased the size of monSTIM1 whilst preserving its functionality. (Kook Y H et al., 2023).
Case study 2
Optogenetic modulation of carotenoid synthesis exemplifies the challenges in reconciling optogenetic control and metabolic efficiency when scaling illumination devices. Independent optimization of optogenetic induction and product yield in each system by the researchers preceded the combination into light-responsive strains by streamlining development. Their approach serves as a principle for designing strains and photobioreactors amenable to industrial-scale optogenetic production.
Figure 2. The EL222 photowired system responds to blue light, which activates genes in the retina under the control of the pC120 promoter. The researchers optimized the culture conditions so that the constitutive beta-carotene production level varied moderately across devices, ranging from 1,000 to 1,200 μg/gCDW. (Pouzet S et al., 2023).
Q: What are optogenetic tools?
A: Optogenetic tools allow optical control of neural activity using light-sensitive proteins like channelrhodopsins (ChR2), halorhodopsins, and archaerhodopsins.
(1) ChR2 acts as a cation channel that depolarizes neurons upon blue light stimulation, e.g. pAAV-CaMKIIa-hChR2(H134R)-mCherry expresses the variant hChR2(H134R) in excitatory neurons.
(2) Proton pumps like archaerhodopsin (ArchT) in pAAV-GFAP-ArchT-EGFP hyperpolarize neurons by pumping protons out upon yellow light stimulation.
(3) Engineered GPCRs like hM3D(Gq) in pAAV-hSyn-HA-hM3D(Gq)-IRES-mCitrine allow modulation of intracellular signaling.
Q: What factors do you need to consider when designing an optogenetic experiment?
A: (1) Desired excitation or inhibition of neurons
(2) Kinetics of the opsin (millisecond precision with ChR2, steady-state silencing with SSFOs)
(3) Wavelength needed (blue, yellow, red opsins available)
(4) Temporal precision needs (milliseconds to minutes)
(5) Targeted cell type specificity using promoters (CaMKIIa, GFAP, hSyn, CAG)
(6) Localization and expression level needed (viral delivery & recombinases like Cre)
(7) Fluorescent reporter (EYFP, mCherry, GFP, iRFP)
Q: What kinds of light-controllable protein systems are available?
A: (1) Excitatory channelrhodopsins like ChR2
(2) Inhibitory pumps like halorhodopsins, archaerhodopsins
(3) Engineered GPCRs like OptoXRs
(4) Tools for controlling biochemical signaling like Opto-CRAC
We can provide well-characterized variants of these proteins using AAV vectors with flexible promoter and reporting options.
Q: How does one select an optogenetic plasmid?
A: Consider the:
(1) Cell type specificity needed via promoter (CaMKIIa, GFAP, hSyn, CAG)
(2) Type of opsin (excitatory like ChR2, inhibitory like ArchT)
(3) Opsin variant kinetics
(4) Wavelength needed
(5) Fluorescent reporter (EYFP, GFP, mCherry, iRFP)
(6) Genomic elements like WPRE
| Cat.No. | Product Name | Price |
|---|---|---|
| VOT2001 | pAAV-camkIIa-hchr2(h134R)-mcherry | Inquiry |
| VOT2002 | pAAV-hSyn-hChR2(H134R)-EYFP | Inquiry |
| VOT2003 | pAAV-hSyn-hChR2(H134R)-mCherry | Inquiry |
| VOT2004 | pAAV-CaMKII-ChR2-mCherry | Inquiry |
| VOT2005 | pAAV-CamKII-EYFP | Inquiry |
| VOT2006 | AAV-hSyn-HA-hM4D-IRES-mCitrine | Inquiry |
| VOT2008 | pAAV-hSyn-HA-hM3D(Gq)-IRES-mCitrine | Inquiry |
| VOT2009 | pAAV-CAG-ArchT-GFP | Inquiry |
| VOT2010 | pAAV-CaMKIIa-hChR2(H134R)-EYFP | Inquiry |
| VOT2011 | pAAV-CaMKIIa-hChR2(E123A)-mCherry | Inquiry |
| VOT2012 | pAAV-TRE-chR2-EYFP | Inquiry |
| VOT2013 | pAAV-CaMKIIa-eArch 3.0-EYFP | Inquiry |
| VOT2014 | pAAV-CaMKII-hChR2 (E123A)-mCherry-WPRE | Inquiry |
| VOT2016 | pAAV-CaMKIIa-mCherry | Inquiry |
| VOT2017 | pAAV-GFAP-hChR2(H134R)-mcherry | Inquiry |
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