WiSciFest 2019: A Retrospective

Posted on by Julia Nepper

This past weekend was the 9th Annual Wisconsin Science Festival, and we at Promega were excited to join in the celebration of science throughout the state. We participated in the Discovery Expo on Thursday and Friday, where dozens of demonstrations and exhibits were scattered throughout the Wisconsin Institute for Discovery building. Thousands of children on field trips filled the halls, eager to poke and prod at strange and exciting new things.

At our table, we talked about the science of bioluminescence. With 3D-printed firefly luciferase models in hand, we showed the glow of recombinant luciferase to the incoming children and explained to them how scientists could use bioluminescence like a tiny “flashlight” to look inside of cells and watch what’s happening. Our learners received a nice little reward for their attentiveness in the form of glow-in-the-dark firefly stickers.

Thank you to the tablecloth for providing the darkness needed to grab this pic.
Continue reading “WiSciFest 2019: A Retrospective”

Pop Quiz: What Scientific Job is Right for You?

Posted on by Becca McKnight

Word Quiz on yellow backgroundI used to love taking magazine quizzes to learn more about myself. I thought it would be fun to create a quiz to help you find out what scientific career path may be the best fit for you. Be open-minded while taking the quiz and remember that this is just for fun!

1. My greatest strength is:

a)      My artistry
b)      My perseverance
c)      My attention to detail
d)      My problem solving skills
e)      My personality- I get along with everyone

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Discussing the Future of Gene Editing at CRISPRcon Midwest

Posted on by Mariel Mohns
crisprcon_banner
Walking in to the first session at CRISPRcon Midwest.

Last week, a diverse group of stakeholders attended CRISPRcon Midwest, hosted by the Keystone Policy Center and the University of Wisconsin–Madison. The goal of the day-long conference was to emphasize the importance and value of gene editing technology, and how it must be communicated deliberately between scientists, the public, policymakers, and other stakeholders.

Julie Shapiro, Senior Policy Director of Keystone Policy Center, acted as Emcee for the event. Given the diverse group of attendees, she mentioned in her opening remarks that the event organizers were “seeking conversation, not consensus” and emphasized the “power of respectful dialogue.” A slide overhead showcased the ground rules for the day, which included statements such as “dare to listen, dare to share, and dare to disagree.”

crisprcon_poll_word_cloud
Word cloud generated from live polling results at CRISPRcon Midwest.

CRISPRcon aimed to included voices beyond those represented by keynote speakers and panelists, so they incorporated live polling through an online app to keep the audience engaged and an active participant in the conversations throughout the day. From the opening remarks, it was clear that this conference would not just deliver on its promise of thoughtful conversation about the science, but build further understanding about the societal impacts of a rapidly advancing technology.

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Promega Receives Wisconsin 75 Sustainability Award

Posted on by Kari Kenefick

There is nothing like a bit of recognition to energize your efforts, right? Promega was recently awarded the 2019 Distinguished Performer: Sustainability award, as one of the Deloitte Wisconsin 75 awardees.

This award is not so much a feather in our cap, as fuel for our sustainability fire both in Madison, and globally. Here are a few details on the award and why Promega was chosen.

The Deloitte Wisconsin 75 awards recognize private sector companies that:

Promega Germany GmbH office.
New Promega Germany (GmbH) office.
  • Are headquartered in Wisconsin
  • Have a majority (>50%) of ownership held by an individual, family or employees stockholders
  • Have at least $50M in annual sales
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Looking Back: Cell-Free Expression Systems Helped to Characterize Proteins Involved in Hypoxia Response

Posted on by Gary Kobs
Structur of a HIF-1a-pVHL-ElonginB-ElonginC complex
Structure of a HIF-1a-pVHL-ElonginB-ElonginC complex

William G. Kaelin Jr., Sir Peter J. Ratcliffe and Gregg L. Semenza were awarded the 2019 Nobel Prize in Physiology or Medicine for their discoveries of how cells sense and adapt to oxygen availability.

Kaelin and Ratcliffe’s labs focused their efforts on the transcription factor HIF (hypoxia-inducible factor). This transcription factor is critical in the cellular adaptation of to changes in oxygen availability.

When oxygen levels are elevated cells contain very little HIF. Ubiquitin is added to the HIF protein via the VHL complex and it is degraded in the proteasome.  When oxygen levels are low (hypoxia) the amount of HIF increases.

In 2001 both groups published articles characterizing the interaction between VHL and HIF, and these articles were referenced by the Nobel Prize Organization in their press release about this year’s award. (1,2). Both studies demonstrated that under the normal oxygen conditions hydroxylation of proline residue P564 enabled VHL to recognize and bind to HIF.

The use of cell free expression (i.e., TNT Coupled Transcription/Translation System) by both labs was key in the characterization of the VHL:HIF interaction The labs utilized HIF and VHL 35-S labeled proteins generated via the TNT system under both normal or in a hypoxic work station to:

  • Determine the affect of ferrous chloride and cobaltous chloride on the interaction
  • Map the specific region of HIF required for the interaction to occur (556-574)
  • Determine the effect of HIF point mutations on the interaction
  • Use synthetic peptides to block the interaction
  • Conclude that a factor in mammalian cells was necessary for the interaction to occur.

Literature Cited

  1. Ivan, M et al. (2001) HIF Targeted for VHL-Mediated Destruction by Proline Hydroxylation: Implications for O2 Sensing. Science 292: 464–67.
  2. Jaakkola, P. et al. (2001) Targeting of HIF-α to the von Hippel-Lindau Ubiquitylation of Complex by O2– Regulated Prolyl Hydroxylation. Science 202, 468–72 .

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In the Face of Failure, Create Your Own Path

Posted on by Darcia Schweitzer

Do you ever wonder whether you’re on the “right” career path? If you’re in academia, the trajectory you should follow can seem pretty rigid—undergrad degree, PhD, postdoc, PI, and then the elusive tenure. Have you considered that there isn’t a single “correct” path?

The hosts of HelloPhD, Josh Hall and Dan Arneman, interview Promega Science Writer Julia Nepper for their podcast.

That’s the message one of Promega’s Science Writers, Julia Nepper PhD, emphasized when she was interviewed recently on the HelloPhD podcast. The HelloPhD podcast offers advice to help students, postdocs, faculty and scientists navigate the hard questions they face every day related to graduate school and careers in science.

In Episode 121: A Teenager Goes to Grad School, Julia offered her insight on dealing with failure and finding a scientific career path that’s right for you. She also shared her unusual story of starting grad school at age 17 and some of the unique experiences she had along the way that led her to choose a career in scientific communications.

To listen to this podcast and learn more about HelloPhD, click here.

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All You Need is a Tether: Improving Repair Efficiency for CRISPR-Cas9 Gene Editing

Posted on by Sara Klink

Ribonucleoprotein complex with Cas9, guide RNA and donor ssDNA. Copyright Promega Corporation.

With the advent of genome editing using CRISPR-Cas9, researchers have been excited by the possibilities of precisely placed edits in cellular DNA. Any double-stranded break in DNA, like that induced by CRISPR-Cas9, is repaired by one of two pathways: Non-homologous end joining (NHEJ) or homology-directed repair (HDR). Using the NHEJ pathway results in short insertions or deletions (indels) at the break site, so the HDR pathway is preferred. However, the low efficiency of HDR recombination to insert exogenous sequences into the genome hampers its use. There have been many attempts at boosting HDR frequency, but the methods compromise cell growth and behave differently when used with various cell types and gene targets. The strategy employed by the authors of an article in Communications Biology tethered the DNA donor template to Cas9 complexed with the ribonucleoprotein and guide RNA, increasing the local concentration of the donor template at the break site and enhancing homology-directed repair. Continue reading “All You Need is a Tether: Improving Repair Efficiency for CRISPR-Cas9 Gene Editing”

Nucleic Acid from FFPE Samples: Effects of Pre-Analytical Factors on Downstream Success

Posted on by Eric Vincent

Part one of three

Peer-reviewed publications containing data dervived from analysis of nucleic acids isolated from FFPE samples have increased dramatically since 2006.

Formalin Fixed Paraffin Embedded samples (FFPE) have been a mainstay of the pathology lab for over 100 years. Initially FFPE blocks were sectioned, stained with simple dyes and used for studying morphology, but now a variety of biomolecules can be analyzed in these samples. Over the past 10 years we have discovered that there is a treasure trove of genomics data waiting to be unearthed in FFPE tissue. While viral RNAs and miRNA were some of the first molecules found to be present and accessible for analysis starting in the 1990s, improvements to DNA and RNA extraction methods have demonstrated that PCR, qPCR, SNP genotyping, Exome and WGS are possible. This has resulted scientific publications of DNA and RNA data generated from FFPE samples starting in 2006, and today we see immense amounts of data generated from FFPE—with nearly 2000 citations in 2018 reporting sequencing of FFPE samples.

Depending on the type of project, prospective or retrospective, the genomics scientist has an opportunity to affect the probability of success by better understanding the fixation process. The challenge with FFPE is the host of variables that have the potential to negatively affect downstream assays.

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It Takes a Village: Automating Plasmid Purification for iGEM

Posted on by Promega

Today’s blog is guest-written by Wihan Adi, a Master’s student majoring in physics at Justus-Liebig-University in Giessen and team member of iGEM Marburg. Although his background is in nuclear and particle physics, his research interests shifted toward affordable biosensors for point-of-care cancer detection, which is how he ended up doing microbiology for iGEM.

Back in March when the iGEM season had just started, Maurice, a fellow iGEM Marburg team member, told me that he was exchanging emails with Margaretha Schwartz from Promega. Given my background as a physics student, Promega was not a household name for me at the time. “So, are you interested in automating a plasmid purification protocol?” asked Maurice. He told me that Promega was willing to supply the Wizard® MagneSil® Plasmid Purification System for this purpose; that was another name that added to my confusion.

Wihan Adi
2019 iGEM Marburg Team

This year, iGEM Marburg is aiming to establish a fast phototrophic organism as a synthetic biology chassis. For this goal we chose Synechococcus elongatus UTEX 2973, with a reported doubling time of 90 minutes. More specifically, we are creating an easy to use toolbox to empower rapid design testing, including genome engineering tools, self-replicating plasmid systems, natural competence and a Golden Gate-based part library. Our team chose to work on phototrophic organisms because we envision accelerating research in this particular field. (Note: Last year, Marburg’s iGEM project won the Grand Prize!)

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The 30th International Symposium on Human Identification: Elevating DNA Forensics

Posted on by Ken Doyle

Thirty Years of ISHI

30 years of ISHI

In the fall of 1989, a small group of forensic scientists, law enforcement officials and representatives from Promega Corporation gathered in Madison, Wisconsin, for the very first International Symposium on Human Identification (ISHI). At the time, DNA typing was in its infancy and had not yet been validated as a forensic method. The available technology consisted of two methods: detection of restriction fragment length polymorphisms (RFLPs) and variable number of tandem repeats (VNTRs). Promega had developed products based on both analytical methods, which essentially provide a DNA “fingerprint” or profile for each individual tested.

Among the attendees at that first symposium was Tom Callaghan, then a graduate student. That experience made a significant impact on his career path. Last week, at ISHI 30, he presented a session on rapid DNA testing. Dr. Callaghan currently serves as a Senior Biometric Scientist for the FBI. In 1999, he was instrumental in launching the FBI’s Combined DNA Index System (CODIS) and in 2003, he became the first CODIS Unit Chief.

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