Latest Posts

All Posts

GFP Fusion Proteins - Making the Right Connection

Posted by Guest Blogger on Apr 9, 2019 9:13:55 AM

This post was contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology and van Leeuwenhoek Centre for Advanced Microscopy (University of Amsterdam).

Tagging a protein of interest with a fluorescent protein to study its function is one of the most popular applications of fluorescent proteins. These fusion proteins enable the observation of proteins in living cells and organisms. Both components of the chimera are encoded by DNA. Since researchers can generate almost any DNA sequence in the way that they like, the design and engineering of fusion proteins is relatively straightforward. However, generating a fusion while keeping all of the native properties of the protein of interest can be challenging. In this blog I discuss strategies to generate fusion proteins and highlight some aspects of their design. 

Read More >

Topics: Fluorescent Proteins

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.

Read More >

Topics: Fluorescent Proteins

The Fluorescent Vegetables in Aptamer Soup

Posted by Aliyah Weinstein on Jan 29, 2019 9:27:00 AM

If you’re been following Addgene on #souptwitter or if you just enjoy a warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named for soup ingredients! From pSOUP to mCherry, satisfying ingredients reagents permeate the molecular biologist’s kitchen lab - and scientists using aptamers might know this best, as most fluorescent aptamers are named for a fruit or vegetable!

Read More >

Topics: Fluorescent Proteins

Tetbow: Bright Multicolor Labeling for Neuronal Tracing

Posted by Guest Blogger on Jan 24, 2019 9:24:20 AM

This post was contributed by Richi Sakaguchi from Kyoto University, and Marcus N. Leiwe and Takeshi Imai from Kyushu University.

Stochastic multicolor labeling is a powerful solution for discriminating between neurons for light microscopy-based neuronal reconstruction. To achieve stochastic multicolor labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced, stochasticity will result in a combinatorial expression of these three genes with different copy numbers, producing dozens of color hues (Livet et al., 2007; Cai et al., 2013). However, the brightness of Brainbow was inherently low. This is because the stochastic and combinatorial expression of fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level.

Read More >

Topics: Fluorescent Proteins

Fluorescent Biosensors for Measuring Autophagic Flux

Posted by Beth Kenkel on Jan 22, 2019 9:41:01 AM

Autophagy (Greek for “self-eating”) is a process by which cytoplasmic material, including organelles, are targeted to lysosomes for degradation. Autophagy is a dynamic process which involves autophagosome synthesis, delivery of materials to be degraded to the lysosome, and degradation of autophagic substrates inside the lysosome. Historically, methods for studying autophagy focused on counting the number of autophagosomes. This approach, however, has inherent limitations because it turns a dynamic process into a static measurement and it provides limited information about what materials or organelles are being targeted for autophagy. The development of several fluorescent autophagy reporters now allows for the measurement of autophagic flux, or the changes in autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this post is to provide an overview of four autophagy biosensors currently available from Addgene.

Read More >

Topics: Plasmid Technology, Fluorescent Proteins

Click here to subscribe to the Addgene Blog
 
Subscribe

 

Recent Posts