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CRISPR Cheat Sheet

Posted by Tyler Ford on May 31, 2018 10:43:15 AM

At Addgene we periodically have Science Clubs where we present developments in biology research to the whole company with the goal of educating both scientists and nonscientists alike. As part of these presentations, we generally create one page cheat sheets that attendees can use to quickly reference information that they (hopefully) learn at science club. In this post you'll find our CRISPR Cheat Sheet from @megearing's recent science club presentation about genome editing and CRISPR. We hope you find this cheat sheet useful! 

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Topics: Genome Engineering, CRISPR

Genome Engineering, CRISPR, and Research in Singapore: Interview with Wei Leong Chew

Posted by Tyler Ford on Feb 6, 2018 9:00:56 AM

In today’s podcast, we sit down with Wei Leong Chew, a researcher at the Genome Institute of Singapore who recently started his own lab. We discuss some of the joys and difficulties of getting a lab up and running, and learn a little bit about what it was like for Wei Leong to work in George Church’s lab as a graduate student.

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Topics: Genome Engineering, CRISPR, Podcast

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

How to Design Your gRNA for CRISPR Genome Editing

Posted by Guest Blogger on May 3, 2017 11:00:00 AM

This Post was updated on May 3, 2017 with additional information and resources. 

This post was contributed by guest blogger, Addgene Advisory Board member, and Associate Director of the Genetic Perturbation Platform at the Broad Institute, John Doench.

CRISPR technology has made it easier than ever both to engineer specific DNA edits and to perform functional screens to identify genes involved in a phenotype of interest. This blog post will discuss differences between these approaches, as well as provide updates on how best to design gRNAs. You can also find validated gRNAs for your next experiment in Addgene's Validated gRNA Sequence Datatable.

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Topics: Plasmid How To, Genome Engineering, Lab Tips, CRISPR

Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

Posted by Beth Kenkel on Dec 15, 2016 10:57:02 AM

Restriction enzyme cloning is the workhorse of molecular cloning; however, one of its biggest limitations is that sequence modifications can only be made at restriction enzyme cut sites. The lambda red system is an alternative method that can be used for cloning or genome engineering and is based on homologous recombination. It allows for direct modification of DNA within E. coli and is independent of restriction sites. The lambda red system is derived from the lambda red bacteriophage and its use as a genetic engineering tool is frequently called recombineering - short for homologous recombination-mediated genetic engineering.  It can be used to make an assortment of modifications: insertion and deletion of selectable and non-selectable sequences, point mutations or other small base pair changes, and the addition of protein tags. It also has the flexibility to modify the E. coli chromosome, plasmid DNA or BAC DNA. 

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Topics: Genome Engineering, Techniques, Microbiology

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