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CRISPR 101: Targeting RNA with Cas13a (C2c2)

Posted by Joel McDade on Sep 21, 2017 10:07:21 AM

CRISPR, and specifically Cas9 from S. Pyogenes (SpCas9), is truly an exceptional genome engineering tool. It is easy to use, functional in most species, and has many application (see a review of CRISPR applications here). That said, SpCas9 is not the only game in town, and several non-SpCas9 molecules have been characterized. Early research suggests that these molecules may circumvent the limitations associated with SpCas9 (see our blog entitled “Which Cas9 do I choose for my experiment”). A novel protein, Cas13a (previously referred to as C2c2), has several unique properties that make it particularly useful and further expand the CRISPR toolbox. This blog post will cover how Cas13a was identified, the structure and function of Cas13a with a focus on what makes this molecule unique, and the various applications of Cas13a.

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Topics: CRISPR, CRISPR 101

Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**

Posted by Beth Kenkel on Sep 19, 2017 9:20:04 AM

Have you ever wanted to selectively kill a subset of cells in your model system? Turns out that with light-inducible photosensitizers and a quick zap of the proper color light, you can do just that.  Photosensitizing dyes and proteins have been around for awhile (check out this review), but the Bruchez and Tsang labs recently developed a photosensitizer composed of the protein complex complex and the MG-2I-dL5** fluorogen that can be used to ablate cells in culture and in vivo.  Read on to learn more about this killer illumination technique!

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Topics: Plasmid Technology

Some Like it Hot: Thermostable GeoCas9

Posted by Beth Kenkel on Sep 14, 2017 8:40:16 AM

Cas9 is the genome editing tool of choice for a number of model organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish, frogs, some bacteria; but not thermophilic (high heat loving) bacteria. Until recently the only available Cas9 proteins were isolated from mesophilic (medium heat loving) bacteria, such as Streptococcus pyogene’s SpCas9. These Cas9 proteins don’t work well at high temperatures, so to use them in thermophiles, bacteria must be grown at lower temps. This approach only works for facultative thermophiles (high OR medium heat loving), but not obligate thermophiles. However, the recent discovery of GeoCas9 by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR/Cas9 genome editing.

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Topics: Genome Engineering, CRISPR

AAV Vector Quality Control: Going the Extra Mile with NGS

Posted by Karen Guerin on Sep 12, 2017 9:44:59 AM

Reproducible data are key to science, so scientists are used to repeating experiments to confirm their findings. But no scientist wants to repeat an experiment because of poor reagent quality. To make sure our AAV vectors are of the highest quality, we undertake a rigorous quality control process - read on to learn more!

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Topics: Inside Addgene, 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: Podcast

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