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New Tools Enable CRISPRa for Neuroscience Applications

Posted by Guest Blogger on Jul 2, 2019 8:51:40 AM

This post was contributed by Patrick Miller-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center.

During development, complex genetic programs specify and assemble diverse arrays of neurons, forming the neuronal circuits that will later be refined through experience. However, studying the genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating gene expression in the central nervous system (CNS). To address this technological gap, a trio of recent papers describe the development of CRISPR activation (CRISPRa) tools for neuroscience, including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches. Here, we’ll highlight these recent advancements and offer commentary on their application to neuroscience research.

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Topics: CRISPR

Isolating Genomic Regions of Interest with the CRISPR System

Posted by Andrew Hempstead on May 30, 2019 8:44:22 AM

While much of CRISPR research has focused on genome editing, numerous discoveries have been made using the Cas9 nuclease in the absence of genomic alterations. These studies utilize a catalytically inactive form of Cas9 known as dCas9 (Jinek et al., 2012). Like Cas9, dCas9 can bind to a specific DNA sequence via a targeting gRNA. But dCas9 does not cleave the DNA. Much of the research using dCas9 has focused on transcriptional activation using a fusion to a transcriptional activator such as VP64 (Gilbert et al., 2013), or repression of transcription through binding a promoter region to inhibit association of transcriptional activators (Qi et al., 2013). However, the fusion of dCas9 with a protein tag allows for the isolation of a genomic region of interest targeted by a gRNA.

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Topics: CRISPR

What's New in CRISPR - May 2019

Posted by Jennifer Tsang on May 28, 2019 8:49:09 AM

In this quarterly blog series, we’ll highlight a few of the new CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more in-depth, but we hope that this blog series will help you find new CRISPR tools for your research!

This time:

  • Ecoli genome-wide CRISPR inhibition
  • Covalent tethering of DNA template to Cas9
  • SECURE base editors
  • Nme2Cas9
  • CRISPR interference in Candida albicans
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Topics: CRISPR, Other CRISPR Tools

It’s CRISPR Week at Addgene!

Posted by Jennifer Tsang on May 13, 2019 8:01:45 AM

In 2012, we received the first CRISPR plasmid deposit. Now we have over 8,000 CRISPR plasmids in the repository that have been shared over 140,000 times to over 75 countries. That’s a lot of CRISPR! As we’ve shared these tools over the years, the amount of CRISPR educational resources hosted on our website and blog have grown. We even have a CRISPR eBook. This week, we’ll continue to grow the ways we share science by hosting our first ever CRISPR Week on social media.

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Topics: CRISPR

Visualizing Genomic Loci with CRISPR-Sirius

Posted by Jennifer Tsang on Apr 18, 2019 8:53:21 AM

The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without changing them. Thus, it has been widely adapted for CRISPR activation and inhibition experiments. But over the past few years, dCas9 has become a robust visualization tool to study the spatial and temporal arrangement of chromosomes and how these arrangements may affect nuclear processes. The latest of these visualization tools is CRISPR-Sirius, the brightest CRISPR-based tool to date for visualizing genomic loci in living cells.

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Topics: CRISPR, CRISPR Visualize

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