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Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

Posted by Various Addgenies on May 15, 2018 8:43:38 AM

Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.

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Expanding the Optogenetics Toolbox with CRY2clust

Article contributed by Brook Pyhtila

The Won Do Heo lab at the Korea Advanced Institute of Science and Technology (KAIST) and Institute for Basic Science (IBS) has developed another useful optogenetic tool that enables robust and efficient oligomerization of target proteins in response to blue light. This tool, CRY2clust, was created by adding a 9-residue peptide to the C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to blue light, CRY2 undergoes a conformational change that permits it to bind to the CIB1 (cryptochrome-interacting basic-helix-loop-helix) protein. After fusing the CRY2 and CIB1 domains to separate proteins of interest, a researcher can cause them to interact by stimulation with blue light.  Similarly, light can also be used to control homo-oligomerization of a protein fused to CRY2, although some studies have shown that this clustering only happens under certain conditions.

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Topics: Hot Plasmids

Split-BioID: An Improved Method for Studying Protein-Protein Interactions

Posted by Beth Kenkel on May 1, 2018 9:57:26 AM

One way to define a protein’s purpose is by its protein-protein interactions (PPIs). These interactions are often modeled as binary relationships, i.e. protein A interacts with protein B; but proteins are social biomolecules. They can be part of multiple dynamic and overlapping complexes that have distinct functions. Many existing methods for identifying PPIs, such as affinity purification mass spectrometry (AP-MS), lack the ability to specifically identify proteins that interact with a particular protein  complex as opposed to an individual protein. The Bethune Lab has overcome this limitation by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent biotinylation technique BioID. The key advantage of Split-BioID is that it allows for the validation of a binary PPI as well as the identification of additional interacting factors.

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Topics: Plasmid Technology, Hot Plasmids

Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

Posted by Various Addgenies on Apr 23, 2018 10:00:31 AM

Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need. Below you'll find our hot plasmid articles from April 2018. If you'd ever like to write about a recent plasmid deposit please sign up here.

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Topics: Hot Plasmids

AAVs for Genome Editing

Posted by Tyler Ford on Mar 27, 2018 9:32:43 AM

Guest blogger Todd Waldman, Professor at Georgetown University, contributed to this post.

Adeno-associated viruses (AAVs) make fantastic gene delivery vehicles for episomal gene expression and are particularly useful for gene delivery to the nervous system. For many years they have also been used to enhance the efficiency of genome editing. In this post we'll walk through a variety of ways you can use AAVs to improve your genome editing experiments (with and without targeted nucleases).

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Topics: Hot Plasmids, Viral Vectors

New Neuroscience Tool: The SF-iGluSnFr Glutamate Sensor

Posted by Tyler Ford on Mar 8, 2018 9:00:00 AM

In a previous blog post we discussed how fluorescent proteins can be used to construct biosensors, biological tools that monitor processes or detect molecules. Here we’ll be diving into the details surrounding SF-iGluSnFr, a recently upgraded biosensor designed to detect glutamate.

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Topics: Hot Plasmids, Fluorescent Proteins

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