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Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

Posted by Kendall Morgan on Feb 24, 2015 2:50:00 PM

CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation in mind. “We had not managed to create a system to allow us to reliably activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or genes to selectively activating them with the CRISPR system was a large one.  The question for her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that they could be used by people with little bioengineering expertise?

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Topics: Plasmid Technology, CRISPR, pooled libraries

Management for Scientists: Delegating is Key

Posted by Joanne Kamens on Feb 20, 2015 3:30:00 PM

This is the fourth in a 5 part series on Management for scientists. Subscribe to the Addgene Career Advice Posts here.

Once you are responsible for managing others you will only be successful in your role if you become a master at delegating tasks and responsibilities. The manager is not expected to DO all the work she is expected to make sure the work gets done and done well. Involving your team effectively is now your new measure of success. It is imperative that you resist the temptation to “just do it myself” or micromanage you don’t have time for that and you won’t be as productive. For example, the many resources avaliable on the Addgene website didn't come into being through the actions of a single person but hrough the coordinated efforts of many individuals.

Check out Joanne's Reddit AMA

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Topics: Career, Management for Scientists

CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

Posted by Guest Blogger on Feb 18, 2015 10:09:22 AM

The following post was contributed by Daniel Bauer and Matthew Canver of Boston Children’s Hospital and Harvard Medical School. Addgene is proud to present a video reprint of the CRISPR article "Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9" from the Journal of Visualized Experiments (JOVE). The video publication by Stuart Orkin and Daniel Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines. Below Bauer and Canver discuss the motivations behind this research.

 

Using CRISPR/Cas9 for Targeted Genomic Deletions

We were inspired to produce intrachromosomal deletions based on the experiments of Kim and colleagues using zinc finger nucleases to harness non-homologous end joining repair (NHEJ) [1]. Our initial work was with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore the paired double-strand break (DSB) approach at a variety of loci. We were pleasantly surprised by the efficiency of the method. One observation was an inverse relationship between deletion size and frequency [3].

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Topics: Genome Engineering, Lab Tips, CRISPR, Protocols

Plasmids 101: E. coli Strains for Protein Expression

Posted by Julian Taylor-Parker on Feb 10, 2015 10:06:00 AM

In a previous Plasmids 101 blog, we reviewed the salient features of several popular strains of E. coli for DNA propagation. While great for cloning purposes, these E. coli strains are not usually well suited for recombinant protein expression. Many challenges can arise when over-expressing a foreign protein in E. coli. We will review the potential pitfalls of recombinant protein expression and some of the most popular commercial strains designed to avoid them.

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Topics: Lab Tips, Plasmids 101

Management for Scientists: Giving Feedback

Posted by Joanne Kamens on Feb 6, 2015 9:46:00 AM

This is the third in a 5 part series on Management for scientists. Subscribe to the Addgene Career Advice Posts here.

Feedback is help and it should result in authentic assistance for the recipient to become more effective and successful. Giving timely and useful feedback is an absolutely required aspect of being a successful manager. Great bosses tell people where they stand clearly and routinely. They are clear with each employee about what they do well and where they need to improve, and they’re also clear about how the person is doing overall. Employees hate to wonder what you think of their work.

 Check out Joanne's Reddit AMA

 

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Topics: Career, Management for Scientists

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