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The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

Posted by Guest Blogger on Nov 3, 2015 10:30:00 AM

This post was contributed by guest blogger Cameron MacPherson at the Institut Pasteur

CRISPR Software and the Piñata Effect

Two years ago I was a part of a group (Biology of Host-parasite Interactions, Institut Pasteur, Paris) that changed genome editing in the malaria community for the better (Nat. Biotechnol., 2014). Given the timing, it shouldn’t be a surprise that the CRISPR system was involved. Today, that same laboratory enjoys a successful edit rate of over 90% in their work editing the genome of Plasmodium falciparum (the parasite that causes malaria). I attribute their success to technical expertise, thoughtful single guide RNA (sgRNA) design, and the abnormally low GC content of the Plasmodium falciparum genome. To put this last point into perspective, the Plasmodium falciparum genome contains only 0.66 million targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable genome, off-targeting is less of a worry and on-targeting likely more efficient. 

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Topics: Genome Engineering, CRISPR

Cpf1: A New Tool for CRISPR Genome Editing

Posted by Mary Gearing on Oct 14, 2015 10:30:00 AM

This post was updated on Dec 5, 2017.

In 2015, Zetsche et al. added to the CRISPR toolbox with their characterization of two Cpf1 orthologs that display cleavage activity in mammalian cells. Like Cas9 nucleases, Cpf1 family members contain a RuvC-like endonuclease domain, but they lack Cas9’s second HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA rather than the two (tracrRNA and crRNA) needed by Cas9 for cleavage. In certain cases, Cpf1 may be better suited for genome editing than Cas9 - read on to learn more about Cpf1 and check out our CRISPR guide for a refresher on CRISPR/Cas9. 

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Topics: Genome Engineering, CRISPR

CRISPR 101: Mammalian Expression Systems and Delivery Methods

Posted by Nicole Waxmonsky on Sep 24, 2015 10:30:00 AM

This post was updated on Dec 4, 2017.

CRISPR technology has been widely adopted for genome editing purposes because it's cheaper, faster, and easier than prior editing techniques. More and more CRISPR tools are being published each month, making CRISPR a great choice for your next experiment!

In this blog post we’ll provide an overview of some CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods.

 

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Topics: Genome Engineering, CRISPR, CRISPR 101

Genome-wide Screening Using CRISPR

Posted by Joel McDade on Aug 18, 2015 10:30:00 AM

This post was updated on Dec 8, 2017.

What genes are important in your phenotype of interest? Many scientists study diseases for which the underlying genetic cause is not entirely known. Identifying which genes are important for a phenotype can lead to a wealth of additional experiments investigating the role of individual genes or entire pathways in a particular disease process. While CRISPR is certainly not the first means to carry out so-called “forward genetic screening experiments”, it is certainly the most robust. In this blog post, we will discuss how CRISPR libraries are being used to perform genome-wide screens and highlight some of the reagents that have been made publicly available through Addgene. 

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Topics: CRISPR, pooled libraries

Addgene Welcomes John Doench to the Advisory Board

Posted by Tyler Ford on Aug 13, 2015 10:30:00 AM

The Addgene Advisory Board consists of prominent members of the bioscience and tech transfer communities who help guide and promote Addgene in its mission to make research easier. Addgene is excited to announce that we’ve added John G. Doench, PhD to the Advisory Board. John Doench is Associate Director of the Genetic Perturbation Platform at the Broad Institute and has worked with many Addgenies to help improve the understanding, curation, and explanation of our CRISPR resources. We recently sat down with Dr. Doench for an interview to help further introduce him to the Addgene community.

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Topics: Interview, Inside Addgene, CRISPR, Podcast

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