Go fISH! Using in situ Hybridization to Search for Expression of a SARS-CoV-2 Viral Entry Protein

Loss of smell (olfaction) is a commonly reported symptom of COVID-19 infection. Recently, Bilinska, et al. set out to better understand the underlying mechanisms for loss of smell resulting from SARS-CoV-2 infection. In their research, they used in situ hybridization to investigate the expression of TMPRSS2, a SARS-CoV-2 viral entry protein in olfactory epithelium tissues of mice.

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Using Wastewater Surveillance to Track COVID-19 Outbreaks

Today’s blog is written by guest blogger, Sameer Moorji, Director, Applied Markets.

Even as countries are now gradually starting to reopen after lockdown, the COVID-19 pandemic is far from over. Researchers around the world continue to find new ways to monitor, prevent and treat the disease. One new way of monitoring COVID-19 outbreaks relies on a somewhat unexpected source: sewage water.

In March 2020, researchers at the KWR Water Research Institute found the presence of SARS CoV-2 RNA in wastewater samples collected near Schiphol airport in Amsterdam and several other sites in Netherlands. The result came within a week after the first case of COVID-19 in the country was confirmed. This study opened the door to the possibility of using wastewater-based epidemiology to determine population-wide infections of COVID-19.

What is Wastewater-based epidemiology?

Wastewater based epidemiology (WBE), or sewershed surveillance, is an approach using analysis of wastewater to identify presence of biologicals or chemicals relevant for public health monitoring. WBE is not new, as wastewater has previously been used to detect the presence of pharmaceutical or industrial waste, drug entities (including opioid abuse), viruses and potential emergence of super bugs. In fact, several countries have been successful in containing Polio and Hepatitis A outbreaks within their geographic locations.

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Choices for Measuring Luciferase-Tagged Reporter Pseudotyped Viral Particles in Coronavirus Research

Coronavirus (CoV) researchers are working quickly to understand the entry of SARS-CoV-2 into cells. The Spike or S proteins on the surface of a CoV is trimer. The monomer is composed of an S1 and S2 domain. The division of S1 and S2 happens in the virus producing cell through a furin cleavage site between the two domains. The trimer binds to cell surface proteins. In the case of the SARS-CoV, the receptor is angiotensin converting enzyme 2. (ACE2). The MERS-CoV utilizes the cell-surface dipeptidyl peptidase IV protein. SARS-CoV-2 uses ACE2 as well. Internalized S protein goes though a second cleavage by a host cell protease, near the S1/S2 cleavage site called S2′, which leads to a drastic change in conformation thought to facilitate membrane fusion and entry of the virus into the cell (1).  

CDC / Alissa Eckert, MS; Dan Higgins, MAMS

Rather than work directly with the virus, researchers have chosen to make pseudotyped viral particles. Pseudotyped viral particles contain the envelope proteins of a well-known parent virus (e.g., vesicular stomatitis virus) with the native host cell binding protein (e.g., glycoprotein G) exchanged for the host cell binding protein (S protein) of the virus under investigation. The pseudotyped viral particle typically carries a reporter plasmid, most commonly firefly luciferase (FLuc), with the necessary genetic elements to be packaged in the particle. 

To create the pseudotyped viral particle, plasmids or RNA alone are transfected into cells and the pseudotyped viruses work their way through the endoplasmic reticulum and golgi to bud from the cells into the culture medium. The pseudoviruses are used to study the process of viral entry via the exchanged protein from the virus of interest. Entry is monitored through assay of the reporter. The reporter could be a luciferase or a fluorescent protein.   

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38 Years After First Release, RNasin Protects COVID-19 Tests

A protein first purified and sold by Promega almost four decades ago has emerged as a crucial tool in many COVID-19 testing workflows. RNasin® Ribonuclease Inhibitor was first released in 1982, only four years after the company was started. At that time, the entire Promega catalog fit on a single sheet of 8.5 × 11” paper, and RNasin was one of the first products to draw widespread attention to Promega. Today, the demand for this foundational product has skyrocketed as it supports labs responding to the COVID-19 pandemic.

What is RNasin® Ribonuclease Inhibitor?

RNA is notoriously vulnerable to contamination by RNases. These enzymes degrade RNA by breaking the phosphodiester bonds forming the backbone of the molecule. To say that RNases are everywhere is barely an exaggeration – almost every known organism produces some form of RNase, and they’re commonly found in all kinds of biological samples. They’re easily introduced into experimental systems, since even human skin secretes a form of RNase. Once they’re present, it’s very hard to get rid of them. Even an autoclave can’t inactivate RNases; the enzymes will refold and retain much of their original activity.

RNasin® Ribonuclease Inhibitor is a protein that has been shown to inhibit many common contaminating RNases, but without disrupting the activity of enzymes like reverse transcriptase that may be essential to an experiment. It works by binding to the RNase enzyme, prevent it from acting on RNA molecules. This is important for ensuring that RNA samples are intact before performing a complex assay.

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Working from Home: Finding Productivity in this New Normal

During this time of adjusting to a new normal, one of the most difficult things that I have had to get used to is being productive in my own home. Work from home (WFH) days are embraced by some people and not by others. For me, transitioning from working in an office and school setting, to working at-home and completing online courses, has led me on a search for answers about how to get the most out of my day. After creating a productive at-home work environment for me, I wanted to share some of my findings with you.

Here are some of the tips that I have found useful:

Section out a portion of your home for work only.

When I first started working from home, I moved room to room working wherever I felt most comfortable. I soon found this affected my organization and time management, so I started keeping all my work in one area. Now, as I sit here writing this post, I know where all of my work is, and I also know that when I walk out of this area I can ‘power down’ my mind knowing I no longer have to do work.

Power off your electronics when not working.

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A Specific & Sensitive Matter: The Trouble with COVID-19 Antibody Tests

From the beginning of this pandemic, scientists around the world have been working around the clock in pursuit of answers that can effectively combat the SARS-CoV-2 virus. One of hardest things for people to grapple with, is all the unknowns: When will this end? When can I safely visit my friends and family again? What if I have it or had it and I don’t even know it?  

The increased availability of serological testing has helped ease people’s minds about their personal COVID-19 status. From a distance, serological testing may seem like a huge milestone in the marathon that is this pandemic. Unfortunately, many of these tests provide murky and inconsistent results.

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Public-Private Initiative to Increase COVID-19 Testing Capacity by Using Promega Maxwell Instrument in India

This blog is written by guest blogger, Dr Rajnish Bharti, General Manager of Promega Biotech India Pvt Ltd.

As COVID-19 cases accelerate, the country of India has decided to scale up testing capacity to 100,000 tests per day in the coming days.

In a major step to counter the coronavirus crisis, Promega India is supporting government agencies through our automated instruments. The Maxwell® RSC instrument is a compact, automated RNA extraction platform that processes up to 48 samples simultaneously in less than 35 minutes. The automated Promega solution allows laboratories to process up to 400 samples in a typical 8-hour shift.

Scientists in India train on the Maxwell RSC 48
Forensic Science Laboratory-Jaipur and SMS Hospital Jaipur join hands together to use Promega Maxwell® RSC 48 to Increase COVID-19 Testing capacity.
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Targeting IL-6: How A Drug That Helped a 6-Year-Old Beat Cancer Can Save COVID-19 Patients

In 2012, a 6-year-old girl named Emily Whitehead was battling acute lymphoblastic leukemia (ALL), one of the most common cancers in children. Her cancer was stubborn. After 16 months of chemotherapy, the cancer still would not go into remission. There was nothing else the doctors could do, and she was sent home. She was expected to survive only a few more months. Her parents would not give up and enrolled her into a clinical trial of a new immunotherapy treatment called chimeric antigen receptor (CAR) T cell therapy. She was the first pediatric patient in the program.

Doctors took T cells from Emily’s blood and reprogrammed them in a lab. They essentially sent her T cells to boot camp where they are trained to find cancer cells and destroy them. The reprogrammed T cells were then injected back into her body. A week into treatment, she started getting a fever, the first sign that the treatment was working and her reprogrammed T cells were fighting the cancer. But soon, she got very sick. All of the indicators suggested that she had cytokine release syndrome (CRS)—also known as the cytokine storm. This happens when cytokines are released in response to an infection but the process cannot be turned off. The cytokines continue to attract immune cells to the infection site, causing damage to the patient’s own cells and eventually resulting in acute respiratory distress syndrome (ARDS). (Learn more about the cytokine storm in this blog.)

Emily was soon on a ventilator. Tests showed that she had extremely high levels of one particular cytokine: interleukin-6 (IL-6). Desperate to keep her alive, her doctors gave her a known drug that specifically targets IL-6. The results were dramatic. After one single dose, her fever subsided within hours, and she was taken off the ventilator. On May 2nd, 2012, she woke up from an induced coma—it was her 7th birthday. Her doctors said they have never seen a patient that sick get better that quickly.

The drug that saved her life was tocilizumab.

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Getting a PhD in Sweatpants: Guest Blog by Dr. Susanna Harris

Today’s blog is guest-written by Susanna Harris, who recently defended her PhD thesis at the University of North Carolina in Chapel Hill.


I just defended my PhD. Nearly six years of blood, sweat, and tears, most of which were cleaned up with Kimwipes while sitting at my desk in a laboratory facing out towards the UNC Chapel Hill football field. Nearly six years of work, all summed up in a handful of slides. Nearly six years of work, explained to my friends, family, and colleagues – a moment I had dreamed of since the fall of 2014.

What I hadn’t dreamed of? That I would be sitting at my small desk in the corner of my room, with no present audience aside from my snoring dogs. That there would be no dinner celebration that carried into a night of fun along Franklin Street. That, unseen by the viewers of my defense, I would be wearing sweatpants as my name changed from Ms. to Dr. Harris.

Pictured: The audience for Susanna’s thesis defense.

Why did I wear sweatpants when I could have worn literally anything in my closet? Because I think it’s hilarious. I believe this situation will end and we will walk away with memories and lessons learned from an extremely difficult time in the history of the world. I want to walk away with one more ridiculous story to add to a long list of “What even was that?” tales from grad school.

Working towards a PhD is hard at any time; let’s not pretend this pandemic isn’t making things even worse. I was fortunate in many ways that my advisor had already moved our laboratory to a new state in 2019, allowing me to adjust to meeting through webcams and working from home before the pandemic changed the lives of all North Carolinians. This has given me a unique perspective to tease out which problems come from distance working and which are the result of Safer-At-Home orders. Based on my experiences, here are a few tips, tricks, and words of warning.

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Just Keep Swimming: How the Wisdom of a Blue Cartoon Fish Can Inspire Us Amid COVID-19

Today’s blog is written by guest blogger Karen Stakun, Global Brand Manager at Promega.

Wise words from a forgetful blue fish are uniting Promega employees during these trying days. Initiated by our VP of Operations as a rallying call to employees and reinforced through a kind gesture from the Hollywood writer and director who dreamed up the fish, I invite you to join Promega as we “Just Keep Swimming.”

Those words were uttered by Dory, a blue tang with short-term memory loss, in the 2003 animated movie Finding Nemo. Now a classic, it tells the story of Marlin, an overprotective clownfish, who searches the ocean for his missing son Nemo. Dory is his sometimes-unwelcome companion. Desperate to find his son, Marlin grows exhausted and begins to feel defeated, but Dory will not let him give up. Her motivation is simple yet potent. “Just Keep Swimming.”

Setting the Scene

As COVID-19 was emerging in China, Promega began scaling up manufacturing in January to meet the growing global need for testing products. As epidemic became pandemic, and demand quickly became unprecedented, we moved swiftly to increase capacity and add more shifts at our Madison manufacturing facilities, all while ensuring the safety of our employees.

All of this takes dedicated people, especially those on our operations team, working long hours in an atmosphere of global uncertainty. Dedication is in abundance at Promega, as every employee feels a deep commitment to humanity’s struggle against this disease. However, Chuck York, our VP of Operations, says he began seeing the team struggle with the never-ending increases in demand. Despite record product totals, it could be demoralizing for a group that prides itself on always being able to deliver what customers need.

That’s when Chuck recalled one of his family’s favorite movies. “I love the never-give-up aspect of Finding Nemo and in particular the net scene.” Toward the end of the movie, Dory and several other fish find themselves caught in a fishing net. With Nemo’s help, the fish realize they can turn Dory’s mantra into action. They keep swimming together in the same direction and break free of the net.   

“I wanted the team to focus on what we could control, doing all we can each day to keep product flowing. And we were and are doing an outstanding job of that. I also hoped to lighten the mood and bring a smile to peoples’ faces. Our ‘net’ is the ever surging COVID-19 demand, but eventually we will overcome if we just keep swimming.”

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