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Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

Posted by Susanna Bachle on Mar 17, 2020 9:32:55 AM

David Liu’s lab created the first base editor in 2016 (Komor et al., 2016) and since then has been trying to expand their precision editing capabilities. Base editors make specific DNA base changes and consist of a catalytically impaired Cas protein (dCas or Cas nickase) fused to a DNA-modifying enzyme, in this case a deaminase. Base changes from C•G-to-T•A are mediated by cytosine base editors (CBEs) and base changes from A•T-to-G•C are mediated by adenine base editors (ABEs). How does this work? Through molecular biology teamwork. The guide RNA (gRNA) specifies the editing target site on the DNA, the Cas domain directs the modifying enzyme to the target site, and the deaminase induces the DNA base change without a DNA double-strand break. But base editors aren’t perfect. They may be slow, can only target certain sites, or make only a subset of base substitutions. 

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Topics: CRISPR, Base Editing

What's New in CRISPR - March 2020

Posted by Jennifer Tsang on Mar 10, 2020 9:15:00 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 more new CRISPR tools for your research!

This time:

  • Optogenetic repression in yeast
  • Base editing
  • CRISPRi in Caulobacter crescentus
  • Multiplexed gRNA for human pluripotent stem cells
  • CRISPR-based DNA demethylation
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Topics: CRISPR, Other CRISPR Tools

The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

Posted by Guest Blogger on Jan 21, 2020 9:30:00 AM

This post was contributed by the Open Repository of CRISPR Screens.

Imagine you’ve just discovered that your favorite gene was described in a CRISPR screen publication. You see this mentioned in the results section, but you had to dig through the supplemental files to see the actual data. Now you’re wondering if your favorite gene is included in other CRISPR screens and whether that data is useful for your research. Fortunately, there’s a growing open repository of CRISPR screen data that might be able to help you out.

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Topics: CRISPR, Other CRISPR Tools

What's New in CRISPR - December 2019

Posted by Jennifer Tsang on Dec 17, 2019 9:37:19 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 (such as these recent blog posts on prime editing, IgnaviCas9, and Nanoblades), but we hope that this blog series will help you find more new CRISPR tools for your research!

This time:

  • CRISPRi in Burkholderia
  • Base editors
  • CRISPR toolkit for epitope tagging
  • Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery
  • Cytosine deaminase for RNA editing
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Topics: CRISPR, Other CRISPR Tools

Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

Posted by Christina Mork on Nov 12, 2019 9:00:00 AM

In 2008 the Quake Lab at Stanford University became interested in exploring biological dark matter – large tracts of the microbial tree of life that remained unexplored. Using new single-cell sequencing approaches, the lab was able to eliminate the need for axenic (pure) laboratory cultures to study these microbes. From 16S rRNA sequencing, hot springs were known to be diversity hotspots containing abundant biological dark matter and so the lab organized a sampling trip to Yellowstone National Park (YNP).

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Topics: CRISPR, Cas Proteins

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