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How to Design Your gRNA for CRISPR Genome Editing

Posted by Guest Blogger on Sep 24, 2020 9:15:00 AM

Originally published May 3, 2017 and last updated Sep 24, 2020

This post was contributed by guest blogger, Addgene Advisory Board member, and Institute Scientist 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, and 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. A more extended discussion of these subjects can be found in two recent review articles (Doench et al., 2017, and Hanna et al., 2020) and references therein.

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Topics: CRISPR, CRISPR gRNAs

Finding Your Science Policy Path

Posted by Guest Blogger on Sep 22, 2020 9:15:00 AM

This post was contributed by Steph Guerra, a AAAS Science and Technology Policy Fellow at the Veterans Health Administration.

“But, seriously, what even is science policy?” 

I have been asked this many times throughout my short science policy career and this seemingly simple question is a moving target. There are a multitude of options for achieving success in a science policy career with many opportunities to pivot and grow along the way. That’s what makes it so wonderful.

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Topics: Science Careers, Science Career Options

Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

Posted by Guest Blogger on Aug 27, 2020 9:15:00 AM

This post was contributed by Patrick Miller-Rhodes from the University of Rochester Medical Center. 

You’ve probably heard of Forster Resonance Energy Transfer (FRET). Through the non-radiative transfer for energy between neighboring fluorophores, FRET can be used to detect the inter- and intramolecular interactions that underlie protein function. However, FRET experiments can be difficult to implement in practice because FRET depends on a number of hard-to-achieve factors. For example, FRET requires that fusion proteins be in close proximity and present in large enough quantities (and the correct stoichiometric ratios) to generate useable data. What’s more, measuring and quantifying FRET is often easier said than done.

Fortunately, a complementary method exists for visualizing protein-protein interactions (PPIs): Bimolecular Fluorescence Complementation (BiFC).

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Topics: Fluorescent Proteins, FRET

The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

Posted by Guest Blogger on Jul 16, 2020 9:15:00 AM

This post was contributed by Max W. Shen from MIT, Alvin Hsu Harvard University, and David R. Liu from the Broad Institute and Harvard University.

Over the course of the last six months, COVID-19 has had a tremendous impact on our world -- as of June 4, 2020, COVID-19 has caused an estimated total of 380,000 deaths worldwide (statistics provided by Google on 6/4/20). Many countries entered lockdown for weeks to months, pausing or terminating employment for a significant fraction of the workforce.

Wet lab scientists are no exception to this effect -- meticulously designed experiments in labs around the world were put on hold indefinitely due to COVID-19. In our lab, we were interested in examining the effect of COVID-19 on worldwide scientific activity. However, this type of open-ended question is difficult to answer quantitatively without broad, unbiased data. Therefore, we chose to examine a dataset of Liu lab plasmid requests from Addgene, as a proxy for global activity in our particular scientific subfields. 

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Topics: Other, COVID-19

SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

Posted by Guest Blogger on May 5, 2020 9:15:00 AM

This post was contributed by Shravanti Suresh from Iowa State University.

Since its appearance, SARS-CoV-2 has spread to almost every part of the world manifesting as a full-fledged pandemic. Containing the spread of this virus has become an utmost priority for countries around the world and to do so, the WHO recommends one strategy: testing, tracking, and social distancing.

With countries like South Korea, one of the earlier epicenters of the outbreak ultimately flattening the curve, it has become evident that widespread testing is crucial in controlling this pandemic. Currently, the CDC uses RT-qPCR tests to diagnose COVID-19 and some serological tests to determine past exposure. However, the limited availability of reagents and equipment and the long turnaround times, have led researchers to turn to other technologies like CRISPR.

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Topics: CRISPR, Cas Proteins, COVID-19

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