Light-activated gene expression has been employed in diverse research areas for decades. More recently, the development of induced pluripotent stem cells (iPSC) and the enhanced flexibility it has brought to in vitro assays has focused attention on in vitro molecular biology approaches. As an example, the Zhang lab at Harvard University developed Light-inducible Transcription Effectors (LITEs) that promote targeted gene expression when exposed to light. Research, like this Nature 2013 article, could be significantly accelerated with Lumos.
Today’s sophisticated biological research requires new tools to assert control over complex cell processes and behavior. Find out how Lumos' throughput, flexibility, and control provide new avenues for in vitro research.
The precise control optogenetics provides offers benefits for the assessment of biochemical and intracellular signalling pathways as well. To date, many cellular pathways and functions have been studied incorporating optogenetic control including the MAPK, and PI3K pathways, Rho family GTPase activation, apoptosis, and protein trafficking. For all of these and more, Lumos provides a flexible, high-throughput platform such that any pathway or interaction can be precisely controlled for in-depth evaluation.
In this example, the Tucker Lab focused on genetic engineering photosensory domains of plant proteins to control protein interactions and protein activity with light. Research, like this Nature Methods 2010 article, could be significantly accelerated with Lumos.
More recently, beginning in 2015 with publications from the University of Tokyo and Duke University, optogenetics has been used to provide additional control to the gene editing technique CRISPR. Results of these initial experiments concluded that optogenetic control of CRISPR/Cas9 is not only possible, but repeatable and reversible as well.
In this example, Polstein and Gersbach engineered a light-activated CRISPR-Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light. Research, like this Nat Chem Biol 2015 article, could be significantly accelerated with Lumos.
Control cell excitability with light
Optogenetics techniques use genetic targeting to express light-sensitive ion channels (e.g. channelrhodopsin-2, ChR2) in targeted cell populations. When optically stimulated with the opsin-specific activation wavelength, those ion channels are activated, causing depolarization or hyperpolarization of the cell membrane.
Stimulating cell cultures with specific wavelengths of light allows you to activate or silence cell activity in targeted cell populations.
Advantages of optogenetic-mediated control of cell activity include:
Standardize activity to reduce well-to-well and batch-to-batch variability
Study a range of physiologically relevant activities in a single well
Explore activity dependent effects of test compounds
Light-evoked activity is artifact-free
By nature, neural networks are complex biological systems. The Maestro is an invaluable research tool for exploration of neural networks in vitro. Augmenting the Maestro with light control instrumentation opens up new realms for decoding life’s circuitry. For example, optogenetic techniques allow researchers to genetically target and control specific cell types within a neural population. Using Lumos, those cells can be independently stimulated while simultaneously recording of real-time neural activity occurs on the Maestro
New advances in neuroscience applications are in reach through Lumos’ ability to deliver specific light–wavelength, intensity and duration–to multiple wells simultaneously—bringing a high-throughput, flexible benchtop approach to optogenetics.
This control facilitates a deeper understanding of the functional network activity of neurons, and in turn, neurological disease research.
Cardiomyocytes cultured on microelectrode arrays (MEAs) create an accessible platform for studying heart-beats in a dish. Cardiomyocyte assays rely on evaluation of parameters, such as repolarization timing, that are tightly coupled to beat rate. Controlling beat rate allows the user to increase physiological relevance and reduce well-to-well variability. Furthermore, the ability to systemically vary beat rate enables detection of use dependent (i.e. beat rate dependent) drug effects.
In this example, we demonstrate that the relationship between repolarization timing (field potential duration, FPD) and beat rate in hiPSC-cardiomyocytes is not accurately described by Fridericia's or Bazett’s and QT correction formulae.