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Light Up Your Experiments with the Michael Davidson Collection

Posted by Lianna Swanson on Oct 31, 2017 9:22:25 AM

Michael Davidson (1950-2015) dedicated his scientific career to 3 major avenues – mentoring young students and instilling a strong work ethic in them, developing educational resources for microscopy, and building new fluorescent protein tools for the scientific community. Davidson took the fluorescent proteins originally developed by Roger Tsien, a frequent collaborator, and expanded on then to revolutionize the study of cell biology. In 2014, Mike Davidson deposited his plasmid tools with Addgene.

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Topics: Fluorescent Proteins, Generating Fusions

Fluorescent Tagging of Endogenous Genes with SapTrap

Posted by Michelle Cronin on Oct 12, 2017 10:26:13 AM

Since the discovery of GFP over 50 years ago, the growing spectrum of fluorescent proteins (FPs) has been an invaluable resource for studying the organization and function of cellular systems. FPs have been used to track protein localization, cell structure, intracellular trafficking, and protein turnover rates. Additionally, by engineering FP fusions associated with cellular organelles, scientists have been able to study many cellular processes, including mitosis, mitochondrial fission/fusion, nuclear import, and neuronal trafficking. Although FPs have enabled discovery of many cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently tagged proteins can lead to improper protein localization, protein aggregation, or disruption of normal protein function, and ultimately misinterpretation of the protein’s cellular role.

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Topics: Fluorescent Proteins, Generating Fusions

Plasmids 101: SunTag and Fluorescent Imaging

Posted by Mary Gearing on Mar 28, 2017 10:30:00 AM

Quick Announcement from the Plasmids 101 Team: In preparation for the release of Addgene's Fluorescent Protein eBook - our next couple of plasmids 101 posts will gain a healthy, fluorescent glow. Stay tuned for more fluorescence-based Plasmid 101 posts in the coming weeks!

In biology as in life, more is often better. More transcription factor binding sites in a promoter lead to higher transcriptional activation. Multiple nuclear localization signals (NLS) increase protein import into the nucleus. In developing their SunTag technology, the Vale and Weissman labs took this biological lesson and created a system to amplify fluorescent signals. Named for the "stellar explosion SUperNova," SunTag can help you turn up the brightness in your fluorescent imaging experiments.

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Topics: Fluorescent Proteins, Generating Fusions

MXS Chaining

Posted by Leila Haery on Feb 7, 2017 10:30:00 AM

High-throughput cloning, in a nutshell, is the systematic combination of different genetic sequences into plasmid DNA. In high throughput cloning techniques, although the specific sequences of the genetic elements may differ (e.g., a set of various mammalian promoters), the same cloning procedure can be used to incorporate each element into the final construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many biological fields. In synthetic biology for example, high-throughput cloning can be used to combine the functions of different genetic elements to generate non-natural tools such as novel biological circuits or sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging technologies, the combination of multiple fluorescent protein sequences to develop diverse fluorescent reporters is a useful application of high-throughput cloning. MXS Chaining is one such technique and has been used to produce complex fluorescent reporter constructs. These fluorescent reporters can be used to detect structure and protein localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu, 2015).

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Topics: Fluorescent Proteins, Generating Fusions

The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

Posted by Kendall Morgan on Jul 29, 2014 2:16:00 PM

In the world of fluorescent proteins and their use for imaging cell biology, Michael Davidson’s lab at Florida State University has been the go-to place. In 2012, his National High Magnetic Field Lab worked with an impressive 1,350 scientists from more than 275 institutions all over the world. In the course of all those collaborations over the years, he and his colleagues built a Molecular Expressions collection including some 3,300 plasmids along with image galleries and educational resources to go with them. This collection of plasmids is available in an easily searchable format on Addgene’s Michael Davidson Fluorescent Protein Collection webpage. (Check back regularly as new plasmids are being added to the webpage every week.)

Over 300 backbones in a rainbow of colors

“It’s a great collection with over 300 backbones alone,” said Addgene’s Lianna Swanson, who has been working with members of the Davidson lab to coordinate the impressively big deposit. “He has every fluorescent protein under the sun, from the standard oldies but goodies (e.g., EGFP and YFP) to the new and improved fruit colors (e.g., apple, papaya, and tomato) and the photoactivatable fluors (e.g. Phamret and Dendra). It’s just such a great collection with such variety.”

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Topics: Fluorescent Proteins, Generating Fusions

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