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Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

Posted by Aliyah Weinstein on Mar 5, 2019 8:08:52 AM

Stochastic multicolor labeling is a popular technique in neuroscience and developmental biology. This type of cell labeling technique involves the introduction of a transgene construct containing fluorescent proteins (XFP) of different colors to label an organ or entire organism. Because each cell can have multiple copies of the transgene that will recombine independently, cells may acquire one of a variety of colors when a combination of XFP are expressed. Each cell remains the same color for its entire lifetime and daughter cells retain the same color, allowing for the fate mapping of cell populations over time. The ability to track single cell dynamics at the organism level has been made possible by tools that allow cells to become persistently fluorescent during development. Stochastic multicolor labeling systems, many based on Brainbow, now exist for a variety of species, cell types, and research applications.

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Topics: Fluorescent Proteins, Cell Tracing, Neuroscience

Neuronal labeling with Spaghetti Monster

Posted by Benoit Giquel on Aug 14, 2018 8:42:01 AM

The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements and behavior. These functions and others are controlled by collections of neurons that are intricately wired into circuits through synaptic connections (Shepherd, 2004). Understanding the structure and function of these neural circuits is essential for neuroscience research. The use of genetic tools for visualizing and perturbing circuits together with the development of methodologies to deliver genes to the CNS have recently made it much easier to map these neuronal networks.

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Topics: Viral Vectors, Neuroscience, Other Viral Vector Tools

Let There Be LITE Plasmids

Posted by Kendall Morgan on Nov 21, 2013 10:56:00 AM

When neuroscience graduate student Silvana Konermann first entered Feng Zhang’s lab at MIT, the use of customizable DNA-binding domains based on transcription-activator-like effectors (TALEs) as anchors for genome engineering applications was still very new. Now, Konermann and her colleagues including Zhang and Mark Brigham have taken the technology to another level with the addition of two light-sensitive ingredients - CRY2 and CIB1 – which they borrowed from Arabidopsis thaliana. The results are light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn them on or off, literally at the flip of a (blue) light switch.

Zhang’s team devised the plasmids now available in Addgene’s repository for use in neurons, both in culture and in living brain tissue. “The brain is such an amazing and dynamic organ,” Konermann explained. “It helps us adapt to influences in our environments. We are able to react and learn. All of this requires genes to be regulated dynamically.”

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Topics: Optogenetics, Viral Vectors, Neuroscience

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