Barking Up the Right Tree: Using NanoLuc to Screen for Canine Distemper Antivirals

Posted on by Natalie Larsen

Canine distemper virus (CDV) is a highly contagious pathogen that is the etiological agent responsible for canine distemper (CD), a systemic disease that affects a broad spectrum of both domestic dogs and wild carnivores. While there are commercially available vaccines for CDV that can provide immunity in vivo and protect canines from contracting CD, there is a strong demand for effective canine distemper antivirals to combat outbreaks. Such drugs remain unavailable to date, largely due to the laborious, time-consuming nature of methods traditionally used for high-throughput drug screening of anti-CDV drugs in vitro. In a recent study published in Frontiers in Veterinary Science, researchers demonstrated a new tool for rapid, high-throughput screening of anti-CDV drugs: a NanoLuc® luciferase-tagged CDV.

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Targeting Glioblastoma Cells by Packaging a Lentiviral Vector Inside a Zika Virus Coat

Posted on by Sara Klink

A recent article published in Cancers demonstrates a new method for targeting glial cells using a lentiviral packaging system that incorporated Zika virus envelope proteins. By using the reporter gene firefly luciferase, researchers demonstrated that a pseudotyped virus could infect cultured glioblastoma cells.

Introduction

Space-fill drawing of the outside of one Zika virus particle, and a cross-section through another as it interacts with a cell. The two main proteins of the viral envelope, the envelope proteins and membrane proteins, are shown in red and purple respectively. The lipid membrane of the envelope is shown in light lavender.The capsid proteins, in orange, are shown interacting with the RNA genome, in yellow, at the center of the virus. The cell-surface receptor proteins are in green, the cytoskeleton in blue, and blood plasma proteins in gold. Drawn by David Goodsell.
Space-fill drawing of the outside of one Zika virus particle, and a cross-section through another as it interacts with a cell. The two main proteins of the viral envelope, the envelope proteins and membrane proteins, are shown in red and purple respectively. The lipid membrane of the envelope is shown in light lavender. The capsid proteins, in orange, are shown interacting with the RNA genome, in yellow, at the center of the virus. The cell-surface receptor proteins are in green, the cytoskeleton in blue, and blood plasma proteins in gold. Drawn and copyright owned by David Goodsell.

Viruses enjoy a fearsome reputation. SARS-CoV-2 is only the latest infectious agent that has garnered attention by becoming a worldwide pandemic. Even the viral name suggests that SARS-CoV-2 was not the first of its type [SARS-CoV is the virus behind the severe acute respiratory syndrome (SARS) that spread worldwide in the early 2000s]. There are many different families of viruses (e.g., coronavirus for SARS-CoV-2 or lentiviruses for HIV-1) and each show a preference to the cell types they want to infect. By investigating the life cycle of viruses to better understand their mechanisms, researchers can discover new opportunities that may be exploited.

In 2015 and 2016, the virus that concerned health authorities was Zika virus (ZIKV). While this virus generally caused mild disease, the babies of women who were infected during pregnancy were at increased risk for microcephaly and other brain defects. These defects were traced back to Zika virus infecting nerve tissue, specifically, glial cells. This discovery provided an opportunity to explore how Zika virus might affect the brain tumor, glioblastoma multiforme (GMB), especially the glioblastoma stem cells (GSCs) that resist conventional treatment and contribute to the poor prognosis for GMB. Studies suggested that Zika virus infection prolonged survival in animal glioma models and selectively killed GSC with minimal effects on normal cells. In fact, the molecules used by ZIKV to enter cells were predominantly found on tumors, not normal cells. Knowing that the ZIKV envelope proteins prM and E provide the target specificity for glial cells, Kretchmer et al. wanted to explore if ZIKV envelope proteins substituted in lentivirus packaging systems would be able to enter glioblastoma cells.

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Ramping Up COVID-19 Testing with the Maxwell® HT Viral TNA Kit

Posted on by Jordan Villanueva
COVID-19 testing with Maxwell HT

John Longshore admits that he was not a big Promega customer before the COVID-19 pandemic. His team uses a wide variety of suppliers to assemble the types of testing protocols needed to serve over 50 hospitals. However, when he began to face supply chain disruptions in early 2020, he needed a supplier he could depend on to support the rapid scale-up of COVID-19 testing, and Promega rose to the occasion.

“When we started working with Promega for bulk isolation reagents, our ask was, ‘Can you supply us with 15,000 isolation reagents per week?’” John says. “The answer was yes, and we have gotten everything we’ve asked for on the dates that it was promised.”

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Adenoviral Vector Vaccines for COVID-19: A New Hope?

Posted on by Ken Doyle

The global war against the coronavirus that causes COVID-19 rages on, spearheaded by efforts to develop effective and safe vaccines. At the time of writing, over 100 COVID-19 vaccine clinical trials were listed in the clinicaltrials.gov database. Recent attention has focused on mRNA vaccines developed by Pfizer/BioNTech and Moderna. If licensed, they would become the first mRNA vaccines for human use.

Other vaccine development efforts are relying on more conventional techniques—using an adenoviral vector to deliver a DNA molecule that encodes the SARS-CoV-2 spike (S) protein. Examples of these adenoviral vector vaccines include the vaccines from Oxford University/AstraZeneca (the UK), Cansino Biologics (China), Sputnik V (Russia) and Janssen Pharmaceuticals/Johnson & Johnson (the Netherlands and USA).

sars-cov-2 coronavirus covid-19 infection with antibodies from a vaccine attacking the virus; several vaccines are in development including adenoviral vector vaccines
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Automation Helps A Graduate Student Monitor COVID-19 With Wastewater

Posted on by Johanna Lee
Kasia Slipko (middle) and her lab at Vienna University of Technology. She and colleagues are exploring using wastewater to monitor viral disease outbreaks.
Kasia Slipko (middle) and her lab at Vienna University of Technology. She and colleagues are exploring using wastewater to monitor viral disease outbreaks.

When Kasia Slipko started graduate school at Vienna University of Technology, Institute for Water Quality and Resource Management, she was interested in studying antibiotic resistant microbes in wastewater. For three years, she evaluated different wastewater treatment methods to find out how to remove antibiotic resistant bacteria. But in the spring of 2020, her research took an unexpected turn. That was when the COVID-19 global pandemic hit, caused by the rapid spread of the SARS-CoV-2 virus. Kasia soon found herself at the forefront of another exciting field: using wastewater to monitor viral disease outbreaks.

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Adapting Holiday Traditions: COVID, Customs and Cookies at Promega

Posted on by Karen Burkhartzmeyer

Adaptation: In biology and ecology, the process or state of adjusting or changing to become more suited to an environment.    

Holiday traditions are certainly taking new forms this year as we all determine how to safely celebrate during a pandemic. It goes without saying that it’s been a tough year. Customs and rituals, large and small, bring peace and comfort. We need those more than ever now, so the challenge becomes finding new ways to honor valued traditions.

Chuck York, VP Manufacturing delivers individually packaged cookies to R&D Scientists on the Promega Madison Campus, adapting this holiday tradition to the life during a pandemic.
This year’s cookie delivery happened with a twist. Chuck York, VP Manufacturing delivers individually packaged cookies to R&D Scientists on the Promega Madison Campus.

Today, we would like to share how one dearly held Promega Madison tradition was able to endure in our COVID-19 world. Adaptation is key. And butter and sugar help, too.

Elaine Day

Promega employees this week were surprised and deeply moved to find that their beloved “Elaine Day” had not become yet another casualty of the pandemic.

“This has been such a difficult year,” says Senior QA Scientist Sue Wigdal. “I had assumed, sadly, that Elaine Day would be cancelled, but to be able to have it and all the thoughtfulness and deliciousness that it brings, was amazing.”  

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SARS-CoV-2 Nucleocapsid Protein and PA28γ: A Role in Pathogenesis?

Posted on by Gary Kobs

The SARS-CoV-2 nucleocapsid protein accounts for the largest proportion of viral structural proteins and is the most abundant protein in infected cells. Nucleocapsid proteins have the job of “packaging” the viral nucleic acid (in this case, RNA). Viral nucleocapsid proteins can also enter the host nucleus and interact with a variety of host proteins to interfere with critical processes of the host cell, including protein degradation. Here we review a study that used an in vitro protein degradation assay to investigate the interaction of the SARS-CoV-2 nucleocapsid protein and the proteasome activator PA28γ.

SARS-CoV-2 structural diagram, showing the SARS-CoV-2 nucleocapsid protein composed of RNA and N protein.

In SARS-CoV-2 infections, the nucleocapsid protein is critical for infection, replication and packaging. The SARS-CoV-2 nucleocapsid protein is not only localized in the cytosol of the host cell but also is translocated into the nucleus. There, it interacts with various cellular proteins that modulate cellular functions, such as the degradation of unneeded or damaged proteins by proteolysis. Researchers have proposed that the protein degradation system plays an important part in coronavirus infection (1).

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How A New SARS-CoV-2 Wastewater Testing Kit is Helping Campuses Reopen

Posted on by Johanna Lee

The fall of 2020 was like no other, especially for universities. The COVID-19 pandemic hit most of the world in the spring, forcing schools and businesses to close. For months, people worked from home and schools switched to online classes. When fall came, universities had a difficult decision to make. Do they have students and staff come back to campus for in-person classes? With students living together in close proximity in dormitories, an outbreak could quickly get out of hand. How can the university monitor and control the spread of the virus to ensure everyone’s safety?

This was when Robert Brooks started getting calls. He’s the Technical Director and Operations Manager at Microbac Laboratories in Oak Ridge, Tennessee. Microbac is a network of privately owned laboratories that provide testing services for food products, environmental samples and the life science industry. Robert has been in the lab industry for 25 years and has established a reputation for taking on difficult problems. “We really try to go that extra mile to help clients solve their issues. That has made a name for us out there. When people have odd-ball issues, they give us a call cause we’re going to take a look at it from a couple different viewpoints and take a step-by-step approach,” he says.

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2020 Promega Award for Biochemistry Recognizes Viral Research, Protein Engineering

Posted on by Jordan Villanueva
Promega Award for Biochemistry image

The 2020 Promega Award for Biochemistry ceremony was a bit different this year. Promega Beijing typically announces the award recipients in a ceremony at the biannual meeting of the Chinese Society of Biochemistry and Molecular Biology (CSBMB). As a result of the COVID-19 pandemic, the 2020 conference was moved online. Despite the unusual circumstances, Promega Beijing held a virtual ceremony to grant the award to Dr. Peng Chen and Dr. Haitao Yang.

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Galloping to Greatness: Meet Kurt the First Cloned Przewalski’s Horse

Posted on by Natalie Larsen

On August 6, 2020, the first successfully cloned Przewalski’s horse was born at the Texas-based veterinary facility, Timber Creek Veterinary, along with a new hope for restoring some much-needed genetic diversity to the species. The successful birth of this foal is the culmination of the collaborative efforts between Revive & Restore, San Diego Zoo Global (SDZG), and ViaGen Equine, and lays the groundwork as an important model for future conservation efforts.

Kurt the first cloned Przewalsk'si horse
Kurt at Timber Creek Veterinary, 09/28/20.
Photo by Scott Stine.

The new Przewalski’s foal (pronounced “shuh-VAL-skees”) has been affectionately dubbed Kurt, in honor of noted animal conservationist, geneticist and pathologist, Dr. Kurt Benirschke. Dr. Benirschke played an instrumental role in founding the Frozen Zoo®, a genetic library comprised of cryopreserved cell lines of endangered species. Established in the 1970s, this collection was built on a foundation of prescient hope, banking on the future development of reproductive and cloning technologies that did not yet exist.

Now thanks to his foresight, that gamble is paying off and the fruits of that labor are literally being brought to life almost 50 years later through Kurt the foal, who is as adorable as he is important to the future of his kind.

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