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Experimenting with New Careers while in Grad School

Posted by Guest Blogger on Feb 25, 2016 10:30:00 AM

Thie post was contributed by guest blogger Melanie Fox, founder and executive director of Central Indiana Science Outreach and a Postdoc at Indiana University School of Medicine.

It’s not always easy to figure out what you want to do after graduate school, at least not while you’re still in the thick of it. About three or four years into my PhD in Molecular Biology, I realized I wanted a career in science outreach: engaging the public to promote an awareness and understanding of science. Like most science PhD programs, mine was geared toward careers in academia or industry. Luckily, I discovered that there are many ways to get a taste of a variety of careers while still working towards your degree. For me, experimenting with the career and networking opportunities available to me as a graduate student culminated in my founding a nonprofit called Central Indiana Science Outreach, or CINSO. We organize fun science events for adults and professional development opportunities for researchers interested in connecting with the public. My experiences in grad school, though often called “nontraditional,” helped prepare me to start CINSO. Here, I share some of the tips I learned along the way and hope they’ll help you make the most of the opportunities you’re provided throughout your PhD program.

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Topics: Career, Career Readiness

CRISPR-Cas9: Tips for Optimizing sgRNA Activity

Posted by Guest Blogger on Feb 19, 2016 10:18:31 AM

 This post was contributed by John Doench of the Broad Institute.

For more infomation on gRNA design, see our post: How to Design Your gRNA for CRISPR Genome Editing

Whether designing a small number of sgRNAs for a gene of interest, or an entire library of sgRNAs to cover a genome, the ease of programing the CRISPR system presents an embarrassment of riches of potential sgRNAs. How to decide between them? By taking into account both on-target efficacy and the potential for off-target activity, experiments utilizing CRISPR technology can provide a straightforward means of determining loss-of-function phenotypes for any gene of interest.

Predicting sgRNA Efficacy

We have recently examined sequence features that enhance on-target activity of sgRNAs by creating all possible sgRNAs for a panel of genes and assessing, by flow cytometry, which sequences led to complete protein knockout (1).

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Topics: Plasmid Technology, CRISPR

Building Global Connections with the International Mentorship Program USA-EUROPE

Posted by Guest Blogger on Nov 6, 2015 10:30:00 AM

This post was contributed by guest bloggers and IMP organizers Rosario F. Godino, Rocío López-Diego, & Zafira Castano Corsino.

When someone says “internationalization” young hearts often shake with the fear of uncertainty. However, internationalization, the ability to cross country boundaries for both professional and personal development, is essential to excelling in the modern world. Talent knows no boundaries and should not be confined by birth country or financial means. Instead, talent should be realized through equal opportunities for all regardless of origin. It is therefore essential that internationalization become a reality available to all youth, our future, around the world.

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Topics: Career, Networking, Career Readiness

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

Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

Posted by Guest Blogger on Oct 27, 2015 11:00:00 AM

 This post was contributed by guest bloggers Erik L. Snapp and Lindsey M. Costantini.

"You underestimate the power of the Dark Side."

--Darth Vader in "Return of the Jedi"

While Vader was referring to the evil side of a mystical "Force," this quote is equally applicable to many microscopy experiments with fluorescent proteins (FPs) localized to compartments other than the cytoplasm. That is, unfortunately, some investigators realize too late that they have missed the impact of dark, non-fluorescent, and misfolded FP-fusions on quantitative imaging experiments and cell physiology in general.

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Topics: Imaging, Fluorescent Proteins

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