Chapter title |
Optogenetics
|
---|---|
Chapter number | 7 |
Book title |
Optogenetics
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3512-3_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3510-9, 978-1-4939-3512-3
|
Authors |
Schumacher, Charlotte Helene, Körschen, Heinz G, Nicol, Christopher, Gasser, Carlos, Seifert, Reinhard, Schwärzel, Martin, Möglich, Andreas, Charlotte Helene Schumacher, Heinz G. Körschen, Christopher Nicol, Carlos Gasser, Reinhard Seifert, Martin Schwärzel, Andreas Möglich |
Editors |
Arash Kianianmomeni |
Abstract |
As a transformative approach in neuroscience and cell biology, optogenetics grants control over manifold cellular events with unprecedented spatiotemporal definition, reversibility, and noninvasiveness. Sensory photoreceptors serve as genetically encoded, light-regulated actuators and hence embody the cornerstone of optogenetics. To expand the scope of optogenetics, ever more naturally occurring photoreceptors are being characterized, and synthetic photoreceptors with customized, light-regulated function are being engineered. Perturbational control over intracellular cyclic-nucleotide-monophosphate (cNMP) levels is achieved via sensory photoreceptors that catalyze the making and breaking of these second messengers in response to light. To facilitate discovery, engineering and quantitative characterization of such light-regulated cNMP actuators, we have developed an efficient fluorometric assay. Both the formation and the hydrolysis of cNMPs are accompanied by proton release which can be quantified with the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF). This assay equally applies to nucleotide cyclases, e.g., blue-light-activated bPAC, and to cNMP phosphodiesterases, e.g., red-light-activated LAPD. Key benefits include potential for parallelization and automation, as well as suitability for both purified enzymes and crude cell lysates. The BCECF assay hence stands to accelerate discovery and characterization of light-regulated actuators of cNMP metabolism. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 11 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Master | 3 | 27% |
Researcher | 3 | 27% |
Student > Ph. D. Student | 2 | 18% |
Student > Doctoral Student | 1 | 9% |
Student > Postgraduate | 1 | 9% |
Other | 0 | 0% |
Unknown | 1 | 9% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 5 | 45% |
Medicine and Dentistry | 2 | 18% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 9% |
Neuroscience | 1 | 9% |
Engineering | 1 | 9% |
Other | 0 | 0% |
Unknown | 1 | 9% |