Cpf1 Update: Comparison to Cas9 and NgAgo

Posted by Mary Gearing on Jul 14, 2016 10:30:00 AM

In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA rather than the two (tracrRNA and crRNA) needed by Cas9 for cleavage. Now, two new studies show that Cpf1 displays lower off-target editing than Cas9, confirming that this protein is well suited for genome editing. 

Find Cpf1 Plasmids at Addgene

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Topics: Genome Engineering, CRISPR

Generating Mouse Models Using CRISPR/Cas9

Posted by Guest Blogger on Jul 12, 2016 10:30:00 AM

This post was contributed by guest bloggers, Wenning Qin and Haoyi Wang.

CRISPR/Cas9 is revolutionizing the mouse gene-targeting field. Mice have long been extremely useful in the lab – they are relatively small and easy to work with, making them the go-to choice for studying mammalian biology. Similar to any model, mice are not without their problems, but much genetic and physiological data have been accumulated over the years using them. Indeed, the future of mouse work is bright as it is now easier than ever to manipulate the mouse genome using CRISPR/Cas9.

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Topics: Genome Engineering, CRISPR

Tips for CRISPR Gene Editing in Mice

Posted by Guest Blogger on Jun 28, 2016 6:59:27 AM

This post was contributed by guest blogger Samantha Young.

The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use in mammalian cells in 2012-2013. Researchers are now using this system in ever more creative ways, (Wang et al., 2013, Cho et al., 2014). There are several variants of the CRISPR/Cas9 system floating around, and many pre-designed plasmids containing these variants ready for purchase. But what is the easiest and fastest way to use the system in mice? We'll have a post that goes into the mouse genome editing process in a bit more detail in the coming weeks, but, in this post, we will outline a simple method for selecting the guide RNA, validating its efficacy in vitro, and using it in mouse embryos to generate gene modified mouse lines. Hopefully this post will help get your in vivo research up and running as soon as possible!

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Topics: Plasmid Technology, Genome Engineering, Lab Tips, CRISPR

Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

Posted by Mary Gearing on Jun 9, 2016 10:30:00 AM

Biologists are going gaga over the newest gene-editing protein - a DNA-cleaving Argonaute from Natronobacterium gregoryi, or NgAgo for short. Addgene has already distributed this plasmid all over the world, and the question on everyone’s minds is: could NgAgo replace CRISPR? Such a drastic shift won’t happen overnight, but there are a few reasons why you might choose NgAgo over CRISPR proteins Cas9 or Cpf1 - keep reading to learn more!

Find New Tips in the Han Lab's NgAgo Protocol Here!

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Topics: Plasmid Technology, Genome Engineering, CRISPR

PiggyBac-ing Through the Genome Editing Field

Posted by Guest Blogger on May 31, 2016 11:30:00 AM

Addgene is proud to announce that we recently acquired the ability to distribute plasmids with the piggyBac™ transposon. These plasmids, when combined with a source of piggyBac™ transposase (available from Transposagen or a licensed distributor) allow you to quickly transfer a DNA sequence from the transposon vector to one of many TTAA sequences distributed throughout the genome. We encourage you to deposit your piggyBac™ transposon vectors with us to help us expand this useful resource. While Addgene cannot distribute plasmids with the piggyBac™ transposase itself, please read on to learn more about this exciting technology from the folks at Transposagen.

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Topics: Plasmid Technology, Genome Engineering

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