Soon after Amanda Capes-Davis started working with CellBank Australia, she received a request from an exasperated graduate student:
This cell line was handed down to me for my project, but I’m getting strange experimental results with the cells.Can you authenticate the cell line?
After performing genetic analyses, Capes-Davis soon had the answer to the student’s experimental woes: the cells did not come from the human tissue type the student was studying. They weren’t even human—they were mouse cells.
“She’d been given this cell line that was behaving differently than expected, and people thought ‘wow, this is an exciting new variant,’ it could tell her more about a particular disease,” Capes-Davis said. “But no, it was a more sinister reason, unfortunately.”
The pandemic caused by SARS-CoV-2 has brought the world to its knees. There have been many deaths, many persons with lingering disease (long COVID) and the inability to vaccinate everyone quickly, for starters. SARS-CoV-2 has not only been a tricky adversary in terms of treatment options to save lives, it’s also been a wily opponent to researchers studying the virus.
Contributing to the existing studies, with their review of the role of inflammasomes in COVID-19, Vora et al. recently published “Inflammasome activation at the crux of severe COVID-19” in Nature Reviews Immunology. In this paper they detail evidence of inflammasome activation and its role in SARS-CoV-2 infections.
Contributions of Those Lost in the SARS-CoV-2 Pandemic I’d like to take a moment to note the uniquely awful nature of the virus at the center of this blog and the paper it reviews. Many of the papers we blog about describe research involving cell lines, mice or another animal model. The closest most reports get to human research subjects is the use of human cells lines. In the Vora et al. report, serum and tissue samples are from actual human patients, some that survived and many that did not survive COVID-19. It’s not lost on us, Dear Reader, the contributions of those that suffered and died due to SARS-CoV-2 infection. Many persons with severe or fatal COVID-19 have made a significant contribution to our understanding of this virus and its treatment options. We owe them, as well as the researchers that have studied SARS-CoV-2, our sincerest gratitude.
Why the Interest in Inflammasomes? For detailed information on inflammasomes you can read Ken’s blog, here. You will find background information there and on our inflammasome web page.
In their paper, Vora et al. provide evidence of inflammasome activation, both direct and indirect, in COVID-19. The authors note:
“Key to inflammation and innate immunity, inflammasomes are large, micrometrescale multiprotein cytosolic complexes that assemble in response to pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) and trigger proinflammatory cytokine release as well as pyroptosis, a proinflammatory lytic cell death.”
The BioPhamaceutical Technology Center Institute (BTC Institute) is a non-profit organization that provides opportunities for people of all ages to learn about life science and biotechnology. This summer, BTC Institute hosted a variety of programs supporting teachers, potential first-generation college students, and many other groups. Each program supports an overall goal to support scientific understanding in our community.
A Celebration of Life: Being Healthy on Earth and In Space
BTC Institute has collaborated with the African American Ethnic Academy in Madison, WI for over 25 years to offer a summer science program for upper elementary and middle school students. This year, A Celebration Of Life XXVIwelcomed 13 students from grades 4-8 every morning for two weeks. Students made ice cream, engineered water filtration devices, and used bioluminescence to learn about preventing the spread of germs. Outside the lab, the students learned tai chi from a Promega employee and toured the Promega culinary garden. Along the way, students learned about historic and contemporary STEM professionals of color associated with each focus area, including astronaut Victor J. Glover and teen entrepreneur Nabil Hamdan.
Lynch syndrome is an inherited condition that significantly increases the risk of developing colorectal and other cancers, often at a young age. People with this condition have close to an 80% chance of developing colorectal cancer in their lifetime. It is the most common form of hereditary colon cancer and causes roughly 3% of all colon cancers. The mutations that cause Lynch syndrome are inherited in an autosomal dominant manner— meaning you only need to have one copy of the gene with a Lynch-associated mutation to be at an increased risk.
It is estimated that 1 in every 279 people have inherited a Lynch-associated mutation (1). Yet despite this prevelence, Lynch syndrome is not well known and ~95% of those with the syndrome don’t know they have it (1).
Lynch Syndrome Cause and Detection
Lynch syndrome is caused by mutations that result in the loss of function of one of the four different major mismatch repair proteins. These proteins act as “proof readers” that correct errors in the DNA sequence that can occur during DNA replication. To determine if Lynch syndrome is likely, simple screening tests can be performed on tumor (cancer) tissue to indicate if more specific genetic testing should be considered. One such screening looks for high levels of microsatellite instability (MSI) in the tumor tissue. High microsatellite instability (MSI-H) in tumor tissue is a functional indication that one or more of the major mismatch repair proteins is not functioning properly.
For those who develop colorectal cancer at an early age or have a family history (immediate family member or multiple family members with colorectal cancer or polyps), screening for Lynch syndrome can offer valuable insight for both patients and their family, as well as for their healthcare provider.
New MSI IVD Test for Colorectal Cancer to Help Identify Lynch Syndrome
The newly released Promega OncoMate™ MSI Dx Analysis System is an FDA-cleared IVD Medical Device and can be used to determine the MSI status of colorectal cancer tumors to aid in identifying those who should be further tested for Lynch syndrome. The OncoMate™ MSI Dx Analysis System builds upon the company’s fifteen year history of supporting global cancer researchers with one of the leading standard tests for MSI status detection. The OncoMate™ MSI Dx Analysis System offers an improved formulation while using the same five markers that have become the gold standard for MSI detection in the research community and is referenced in over 140 peer review publications (2,3).
TheOncoMate™ MSI Dx Analysis System is designed to provide physicians with a functional, molecular measurement of the level of DNA mismatch repair deficiency demonstrated within their patient’s colorectal cancer tumor. MSI testing is recommended to identify candidates for further diagnostic testing for Lynch syndrome. (2–4). The System is part of a broader workflow that includes DNA extraction from FFPE tissue samples, quantitation of DNA, amplification of specific microsatellite markers using multiplex PCR, fragment separation by capillary electrophoresis, and data analysis and interpretation software. The OncoMate™ MSI Dx Analysis System is available in certain countries. Visit the OncoMate™ MSI Dx Analysis System webpage to learn more.
Promega previously announced a CE-marked version of the OncoMate™ MSI Dx Analysis System in France, Germany, Austria, Poland, UK, Ireland, Belgium, Netherlands, Luxembourg, Spain, Italy, Switzerland, Denmark, Sweden and Norway.
On September 4th, 2021 we celebrate National Wildlife Day. This day helps cherish our planet’s biodiversity and recognize issues that impact wildlife. Take a look at three Promega blogs that highlight preservation and conservation efforts being made to support our natural world.
Depression is not simply a mood disorder, a feeling of sadness, or being ill at ease. Depression can completely shut a person down, manifesting as an inability to make decisions, to take action, to think. Even sleep is affected by depression.
Researchers and clinicians who treat depression are learning that the physical manifestations can be mirrored by internal, cellular changes. Some people with depression have decreases in their gray matter volume, particularly in areas like the hippocampus (important to memory, learning, and emotions) and prefrontal cortex (where higher-level thought and planning abilities are based).
Additionally, imaging has shown a decrease in the number of synapses—the structures through which electrical or chemical signals are passed between neurons and other cells—in persons with chronic depression. Without the signals that synapses transmit, brain function is disrupted.
And without intervention in depression, synapse decrease can continue.
While there are drugs and behavioral therapies to treat depression, these therapies can be slow to act and sometimes ineffective. In addition, once synaptic loss has occurred, these therapies are less effective.
“It has long been recognized that these compounds (serotonergic psychedelics like psilocybin) may have therapeutic potential for neuropsychiatric disorders, including depression, obsessive-compulsive disorder and addiction”.
Cell-free gene expression systems are a staple tool for the researcher seeking to understand the regulation of transcription and translation. Many factors can affect the efficiency of cell-free gene expression including vector sequence, reaction components and the template DNA concentration. One factor that has not been extensively studied is how DNA template length influences gene expression.
When we think of viruses, we often think of diseases, pandemics and death. Our impression of viruses is that they are “bad”. But viruses could also be a possible cure for the deadliest disease in modern history: cancer. The therapeutic effects of “good” cancer-killing oncolytic viruses have been documented over a century ago. Records from as early as 1904 described a 42-year old woman with acute leukemia who experienced temporary remission after an influenza infection. Other early reports showed spontaneous remission of Hodgkin lymphoma and Burkitt’s lymphoma after natural infections with the measles virus.
Despite the long history, oncolytic viruses have only recently gained momentum in the scientific community. Dr. Aldo Pourchet, CSO and co-founder of Omios Biologics—a biotech startup in the San Francisco Bay area—is determined to harness the power of oncolytic viruses to develop a new generation of cancer immunotherapy.
How Oncolytic Viruses Work
“One thing that we know for sure is that you need the immune system to fight the cancer,” says Pourchet. “You need to recruit the immune system, and probably the best thing we know for recruiting the immune system is viruses. Our immune system evolved to detect them immediately. That’s why we are still on Earth. It’s because we have been able to fight deadly viruses.”
Today’s guest blog is written by Melissa Martin, a global marketing intern with Promega this summer. She will be a senior this fall at the University of Wisconsin-Madison where she is double majoring in zoology and life sciences communication, with a certificate in environmental studies.
Congrats! You are attending a university and pursuing a challenging, yet rewarding, undergraduate science degree. Getting to this moment probably included lots of late nights spent studying or worrying while applying to your dream college. However, now that you are here you will find that classes provide a lot of information. You can even take your education one step further by getting hands-on experience in a research lab.
Working in a lab is not only about making your resume look good. It offers a real-world experience that directly enhances your learning experience and can even guide your future. For example, your experiences in the lab can teach you basic skills (pipetting, determining concentrations, performing titrations, etc.) that will be useful in a variety of science professions.
Today’s post is written by Michael Curtin, Senior Product Manager, Reporters and Signaling.
Inflammation is a defense mechanism that the body employs in which the immune system recognizes and removes harmful and foreign stimuli and begins the healing process. Inflammation can be either acute or chronic. Chronic inflammation is also referred to as slow, long-term inflammation and can last for prolonged periods (several months to years); chronic inflammation is caused by immune dysregulation. This typically takes the form of the body’s inability to resolve inflammation resulting from overproduction of inflammatory cytokines and chemokines, as well as danger-associated molecular patterns (DAMPs) released from dying cells (2). Tumor Necrosis Factor (TNF) is the primary cytokine involved in many common inflammatory diseases and is where many therapies targeting inflammation are focused.
Recent research that RIP kinases (RIPK1 and RIPK3) are important regulators of innate immunity via their key roles in cell death signaling during cellular stress and following exposure to inflammatory and infectious stimuli. RIPK1 has an important scaffolding role in pro-inflammatory signaling where it interacts with TRADD, TRAF1 TRAF2, and TRAF3 and TRADD can act as an adaptor protein to recruit RIPK1 to the TNFR1 complex in a TNF-dependent process. RIPK1 plays a kinase activity-dependent role in both apoptotic and necroptotic cell death. A review article by Speir et al. (1) discusses the role of RIP kinases in chronic inflammation and the potential of RIPK1 inhibitors as a new therapeutic approach for the treatment of chronic inflammation. RIPK1 or Receptor Interacting Protein Kinase 1 is a serine/threonine kinase that was originally identified as interacting with the cytoplasmic domain of FAS. Promega offers several reagents that make studying RIPK1 easier- these include our RIPK1 Kinase Enzyme Systems which includes RIPK1 (Human, recombinant; amino acids 1-327), myelin basic protein (MBP) substrate, reaction buffer, MnCl2, and DTT and is optimized for use with our ADP-Glo Kinase Assay.
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