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AAVs CREATed for Gene Delivery to the CNS and PNS

Posted by Beth Kenkel on Sep 28, 2017 10:01:35 AM

Adeno-associated viral (AAV) vectors are the most frequently used gene-transfer tools in the study of the brain and spinal cord, which together are known as the central nervous system (CNS). AAVs are popular tools because: 1) their genomes are easy to manipulate, 2) they have long-term expression; and 3) they have limited toxicity. However, a key challenge of using AAVs for neuroscience research is the lack of a method for genetically manipulating neurons throughout the whole brain. Neurons of the peripheral nervous system (PNS), which connect the heart, lung, gut, and other organs to the CNS, are also an important target for gene delivery, especially for the study of pain. While many new capsids (i.e. the part of the virus that determines tropism) have been developed that increase transduction efficiency, none allow for simple and efficient transduction of both the CNS and PNS.That is until the Gradinaru Lab at Caltech stepped up to the challenge.

Find the PHP Plasmids Here!

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Topics: Plasmid Technology, Hot Plasmids, Viral Vectors

Hot Plasmids Podcast Episode 2: New RFPs, AAVs, & More

Posted by Tyler Ford on Sep 8, 2017 3:52:44 PM

In the second episode of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins, AAV tools for targeting the nervous system, and vectors for zebrafish engineering. You can find additional hot plasmids in our quarterly newsletter or on our hot plasmids webpage.

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Topics: Hot Plasmids, Podcast

DIY DNA Ladders from Penn State University

Posted by Beth Kenkel on Jul 14, 2017 10:30:00 AM

Two plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently deposited with Addgene. A team of undergraduate students led by Dr. Song Tan at Penn State developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate 100 bp or 1 kb DNA ladders, respectively. Unlike many commercially available ladders, the 100 bp ladder works well for both agarose and native polyacrylamide gels.

Get Tips on Verifying Your Plasmid

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Topics: Hot Plasmids, Lab Tips

Luminescent Imaging with Nano-lanterns

Posted by Mary Gearing on May 25, 2017 10:30:00 AM

Fluorescent imaging techniques have become indispensable tools for molecular and cell biologists over the last two decades, but their use can be limited by a few caveats. Since fluorescent proteins (FP) require external light activation, you can’t use fluorescence to monitor processes directly affected by light. Long-term light exposure can also lead to cellular phototoxicity, and experimental success can be affected by both autofluorescence and photobleaching. Researchers have long been interested in using luminescence to get around these issues, but this solution wasn’t practical due to the low intensity of luminescent proteins. To make luminescent imaging a reality, Addgene depositor Takeharu Nagai and colleagues at Osaka University have developed Nano-lantern technology. Nano-lanterns contain a Renilla luciferase variant fused to an FP; when supplied with a luciferase substrate, the luciferase transfers energy to the FP, resulting in a fluorescent signal. Since their first publication in 2012, the Nagai laboratory has assembled a collection of multicolored nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion proteins.

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Topics: Hot Plasmids, Fluorescent Proteins

New Podcast Segment: Hot Plasmids

Posted by Tyler Ford on May 17, 2017 11:37:42 AM

We’re breaking into more audio and video on the Addgene Blog and Addgene website. As we push forward with these efforts, you’ll find new ways to learn about science careers, lab protocols, and, of course, plasmids. Today we’re trying a new way to present plasmid info with a new segment on the Addgene podcast - The Hot Plasmids Segment. Click on the player below to listen to a quick (~5 min) Hot Plasmids podcast that introduces you to 4 new plasmid technologies from one of our recent newsletters.

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Topics: Hot Plasmids, Podcast

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