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An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

Posted by Guest Blogger on Jan 27, 2015 10:13:47 AM

This post was contributed by Jordan Ward who is a postdoctoral fellow at UCSF.

Emerging CRISPR/Cas9 editing technologies have transformed the palette of experiments possible in a wide range of organisms and cell lines. In C. elegans, one of the model organisms which I use to study gene regulation during developmental processes, CRISPR/Cas9 allows us to knock out sequences and introduce mutations and epitopes with unprecedented ease. In the last year, several advances in C. elegans genome editing using CRISPR/Cas9 have emerged, which I will describe below. These new C. elegans approaches rapidly enrich for editing events without the need for any selective marker to remain in the edited animal. To my knowledge these approaches have not yet been extended to other organisms/cell lines, though it is likely that many aspects will broadly improve editing efficiency.

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Topics: Plasmid Technology, Genome Engineering, CRISPR

7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

Posted by Guest Blogger on Jan 21, 2015 8:30:00 AM

This post was contributed by Dora Farkas of www.FinishYourThesis.com and is one of many posts in our career series. Click here to subscribe to our career posts.

“The only healthy communication style is assertive communication.” -  Jim Rohn, Author, Entrepreneur, Motivational Speaker

Do you have a difficult supervisor, or do you have to work with difficult people? Each time I ask this question at my workshops, I get nods from nearly every participant.

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Topics: Career

Addgene @ Keystone: Thoughts on Precision Genome Engineering and Synbio

Posted by Eric J. Perkins on Jan 15, 2015 8:50:00 AM

It's been about 14 years since I last attended a Keystone Meeting – far too long. Holding these meetings in relatively isolated resorts creates a sense of comradery with fellow attendees from the moment you arrive. Getting off the plane in Bozeman Sunday night, it was easy to spot meeting participants. They were the ones holding poster tubes (or as our baffled flight attendant called them, "long, skinny things") and generally exuding a very-tired-but-very-excited attitude. Riding up to the resort in the shuttle, our driver regaled us with tales of back country skiing, fly fishing, and local grizzly bear attacks. He described one such recent attack as "hilarious". Welcome to Montana!

Though sadly I will not be attending the entire meeting, Monday's talks alone were worth the trip. Dr. Dana Carroll's excellent keynote address was the first of 19 talks given over the course of the day. His talk, which focused on the history of genome engineering from ZFNs through TALENs and CRISPR-Cas nucleases, provided important context for the rest of the day. He was followed by three of the biggest names in the CRISPR-Cas9 field – Jennifer Doudna, Feng Zhang, and Keith Joung. All Addgene depositors! Addgene was mentioned specifically in Dr. Zhang's introduction. His willingness to share reagents so freely with the academic community has clearly made a huge impact on this field.

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Topics: Genome Engineering, Scientific Sharing, Synthetic Biology, CRISPR

Plasmids 101: Cre-lox

Posted by A Max Juchheim on Jan 13, 2015 10:47:00 AM

In previous posts for our Plasmids 101 series, we examined a number of important plasmid elements – promoters, origins of replication, protein tags, and antibiotic resistance markers (just to name a few). In this edition, we’re going to take a look at a very interesting tool that can be used for creating (excuse the pun) specific, targeted DNA modifications in transgenic animals, embryonic stem cells, and/or tissue-specific cell types: Cre-lox recombination.

What is Cre-lox?

The Cre-lox system is a technology that can be used to induce site-specific recombination events. The system consists of two components derived from the P1 bacteriophage: the Cre recombinase and a loxP recognition site. The P1 bacteriophage uses these components as part of its natural viral lifecycle, and researchers have adapted the components for use in genome manipulation.

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Topics: Plasmid Technology, Plasmid Elements, Plasmids 101

Management for Scientists: Seeking Feedback

Posted by Joanne Kamens on Jan 8, 2015 9:34:00 AM

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

This second installment in the Management for Scientists series will focus on an aspect of communication especially important for a manager – getting feedback from the team. Successful management can almost be boiled down to one, key concept: Creating a culture of excellent, effective communication between all members of a team. As described here in a 2012 Intuit blog post, a study coming out of MITs Human Dynamics Laboratory identified five characteristics of very successful teams.

Check out Joanne's Reddit AMA

Here they are annotated with my comments:

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Topics: Career, Management for Scientists

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