Creative Biolabs

Optogenetics Tools

Optogenetics Tools

Optogenetics refers to "optical stimulation plus genetic engineering", which combines optical and genetic engineering to manipulate cells and their information processes. Optogenetics has been widely used to study the structure, activity, function and cell physiology of non-nervous neural circuits. In 2010, optogenetics was rated as one of the "Technology Achievements in Science and Technology" by "Nature Methods" magazine. This technology can be applied to the treatment of various neurological disorders, such as epilepsy, Parkinson's disease, chronic pain, depression Disease, addiction, etc.

Optogenetics is a technology that integrates multiple disciplines such as optics, software control, gene manipulation technology, and electrophysiology. The main principle is to use gene manipulation technology to transfer optical-sensing genes (such as ChR2, eBR, NaHR3.0, Arch or OptoXR, etc.) into specific types of cells in the nervous system. optical sensitive ion channels will be selective for the passage of cations or anions under different wavelengths of optical stimulation, such as Cl-, Na+, H+, K+, which will cause changes in the membrane potential on both sides of the cell membrane to achieve selective excitement to the cell or the purpose of suppression.

Fig.1 The concepts of different actuators for activation and inhibition of cells. Fig.1 The concepts of different actuators for activation and inhibition of cells.

Optic-sensitive proteins can be used to activate neurons, inhibit neurons and control G protein coupling signals in vivo. The typical optic-sensitive channel protein used to activate neurons is ChR2 (channelrhodopsin). ChR2 is an ion channel protein, which will induce the opening of cation channels after the excitation of blue optic (the maximum excitation peak is around 470nm wavelength), thereby promoting the depolarization of neurons, inducing action potentials, and activating neurons. A typical optic-sensitive protein that inhibits neuronal activity is NpHR (halorhodopsin), which will induce the opening of chloride channels under the excitation of yellow-green optic (the maximum excitation peak is near 590nm wavelength), and the influx of chloride ions causes Hyperpolarization of neurons, thereby inhibiting the generation of neuronal action potentials. In addition, there is Arch (paleopurin), which is found in Salinasodium sphaeroides. Under the excitation of yellow-green optic (the maximum excitation peak is around 566nm wavelength), this protein induces the outflow of protons, thereby generating hyperpolarized signals, and further inhibits the activity of neurons.

Chemogenetics Optogenetics
Resolution
  • Minutes, and CNO acts on neurons
  • Takes 2 hours to remove from the membrane
  • High temporal and spatial resolution.
  • Sub-millisecond and millisecond level in neuron regulation.
  • The spatial accuracy can reach the level of single cell.
  • At the moment the optic stops, the effects of optic stimulation can almost stop at the same time
Experimental Operations
  • Simple operation, only intraperitoneal injection or feeding is enough
  • Compared with optogenetics, no need for craniotomy to embed fiber
The operation is complicated, and it needs to bury the fiber, operate the laser controller and the catheter and other accessories
Experimental Duration
  • Continuous dosing is feasible
  • Suitable for long-term neuronal loop regulation research
  • Due to mechanical limitations such as laser heat generation,
  • Processing neurons for a long time is almost impossible
Cat Product Name Promoter Cell Types Key Components Fluorescent Expression
NTA-2011-ZP51 pAAV-CaMKlla-DIO-hM4D(Gi)-eGFP CaMKlla Glutamatergic Neuron DIO-hM4D(GI) EGFP Inducible Cre expression
NTA-2011-ZP52 pAAV-EF1a-DIO-hM3D(Gq)-mCherry EF1a Broad spectrum DIO-hM3D(Gq) mCherry Inducible Cre expression
NTA-2011-ZP53 pAAV-EF1a-DIO-hM4D(Gi)-mCherry EF1a Broad spectrum hM4D(Gi) mCherry Normal expression
NTA-2011-ZP54 PAAV-GFAP-hM3D(Gq)-mcherry GFAP Astrocyte; Neural Stem Cell hM3D(Gq) mCherry Normal expression
NTA-2011-ZP55 PAAV-GFAP-hM4D(Gi)-mcherry GFAP Astrocyte; Neural Stem Cell hM4D(GI) mCherry Normal expression
NTA-2011-ZP56 pAAV-hSyn-DIO-hM3D(Gq)-mCherry hSyn Neuron DIO-hM3D(Gq) mCherry Inducible Cre expression
NTA-2011-ZP57 pAAV-hSyn-DI0-hM3D(Gq)-eGFP hSyn Neuron DIO-hM3D(Gq) EGFP Inducible Cre expression
NTA-2011-ZP58 pAAV-hSyn-DlO-hM4D(Gi)-mCherry hSyn Neuron DIO-hM4D(Gi) mCherry Inducible Cre expression
NTA-2011-ZP59 pAAV-hSyn-DIO-hM4D(Gi)-EGFP hSyn Neuron DIO-hM4D(Gi) EGFP Inducible Cre expression
NTA-2011-ZP60 pAAV-hSyn-HA-hM3D(Gq)-IRES-mCitrine hSyn Neuron hM3D(Gq) mCitrine Normal expression
NTA-2011-ZP61 pAAV-hSyn-HA-hM4D(GI)-IRES-mCtrine hSyn Neuron hM4D(GI) mCitrine Normal expression
NTA-2011-ZP62 pAAV-hSyn-hM4D(Gi)-mCherry hSyn Neuron hM4D(GI) mCherry Normal expression

How to use optogenetics virus tools for research?

Step1: Find the right optic-sensitive protein

Optic-sensitive proteins can be divided into two types: activation type and inhibitory type, which can cause neuron activation or inhibition. There is a negative correlation between its optic sensitivity and kinetics. The activation or inhibition ability is closely related to the precise control of time. Therefore, finding a suitable optic-sensitive protein according to the different characteristics of the optic-sensitive protein is the first step.

Step2: Introduce optic-sensitive protein genes into target cells

Transfection, virus injection, or transgenic animals are used to input genes encoding optic-sensitive proteins into target cells. Among them, virus injection is the leading method of optogenetic research, mainly using lentivirus and adeno-associated virus.

Step3: Regulate optic signal

The optic can be guided into the study area by introducing optical fiber or controlling the laser, and choosing different parameters (wavelength, optic intensity, frequency and duty cycle) for optic stimulation to achieve time control of neuronal activity; by selective irradiation cell local methods to achieve spatial regulation of neuronal activity.

Step4: Collect the output signal and read the result

Generally, electrodes are used to record the voltage changes inside and outside the cell membrane of neurons, and fluorescent biosensors can be used to detect different cell values. Through methods such as electrophysiological recording or behavioral testing, optic stimulation can present changes in neurons, neural circuits or animal behaviors.

Fig.2 Three primary components in the application of optogenetics. Fig.2 Three primary components in the application of optogenetics.

In addtion to providing neural viral vector tracers, Creative Biolabs also offers relevant AAV packaging services. For the pre-made AAV tools, please contact us for more details.

For Research Use Only. Not For Clinical Use.
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