The Wnt/β-catenin pathway, long studied in the context of developmental biology, has become increasingly recognized for its role in a wide range of human diseases. Its dysregulation has been implicated in cancer, fibrosis, immune modulation, and neurodegenerative conditions—making it a clinically actionable target across diverse therapeutic areas1. In this blog, we cover the fundamentals of Wnt/β-catenin signaling, highlight ongoing research efforts to understand its role in disease, and show how combining live-cell imaging with luminescent assays complements functional studies.
Continue reading “Understanding Wnt Signaling Through β-Catenin Localization in Live Cells”Author: Anna Bennett
Modified Nucleotides in IVT: Small Changes, Big Impact
In our final blog post on double-stranded RNA (dsRNA), we turn our attention to the chemical building blocks of mRNA therapeutics—modified nucleotides. These seemingly minor changes to the RNA sequence play a crucial role in the success of mRNA-based vaccines and treatments. However, they also introduce complexities in accurately detecting and quantifying unwanted dsRNA byproducts— key steps in ensuring the therapeutic efficacy of your mRNA product.
What Are Modified Nucleotides?
Modified nucleotides are ribonucleotides containing chemically altered nucleosides — like specialty ingredients swapped into a classic recipe to improve taste and nutrition. Just as a chef might use a lactose-free milk or gluten-free flour to make a dish easier to digest without changing its core structure, scientists use chemically altered nucleosides during in vitro transcription (IVT) to improve how mRNA therapies perform. These modifications replace their natural counterparts (e.g., uridine or cytidine) in the final RNA product. Their incorporation improves the performance and safety of mRNA therapeutics in several ways:
Continue reading “Modified Nucleotides in IVT: Small Changes, Big Impact “dsRNA QC Considerations: How I Learned to Stop Worrying and Love my IVT Reactions
As mRNA therapeutics continue to expand across clinical pipelines, one persistent challenge remains for developers: reducing double-stranded RNA (dsRNA) contaminants that can compromise safety and efficacy. These unintended byproducts of in vitro transcription (IVT) can trigger unwanted immune responses and reduce the potency of the final product. Developers must prioritize dsRNA detection and control as essential steps in the process. In our previous blog post we offered a high-level discussion of what is double-stranded RNA (dsRNA), its biological function, and importance of detection in a therapeutic context. Here, we’ll take a closer look at origins of dsRNA contamination, quality control measures, and improvement strategies.
Large-scale production of single-stranded RNA (ssRNA) for mRNA-based therapeutics is primarily done through in vitro transcription (IVT), an enzymatic process designed to generate high-yield, functional mRNA transcripts from a DNA template. This process uses purified RNA polymerase enzymes, such as T7, that recognize specific promoter sequences in the DNA template, generating the RNA transcripts of interest. However, IVT reactions also generate unwanted dsRNA byproduct. Below, we delve into some of the major quality control (QC) considerations and strategies to reduce dsRNA byproducts.
Continue reading “dsRNA QC Considerations: How I Learned to Stop Worrying and Love my IVT Reactions”Bioluminescence vs. Fluorescence: Choosing the Right Assay for Your Experiment
From enzyme activity to gene expression, light-based assays have become foundational tools in life science research. Among these, fluorescence and bioluminescence are two of the most widely-used approaches for detecting and quantifying biological events. Both rely on the emission of light, but the mechanisms generating that light—and the practical implications for experimental design—are quite different.
Choosing between a fluorescence or bioluminescence assay isn’t as simple as picking between two reagents off the shelf. Each has strengths and limitations depending on the application, instrumentation, and biological system. In this blog, we’ll walk through how each method works, where they shine (and where they don’t), and what to consider when deciding which approach is right for your experiment.
Continue reading “Bioluminescence vs. Fluorescence: Choosing the Right Assay for Your Experiment “No More Dead Ends: Improving Legionella Testing with Viability qPCR
Legionella is the causative agent of Legionnaires’ disease, a severe form of pneumonia with a mortality rate of around 10%. Contaminated water systems, including cooling towers and hot water systems, serve as primary reservoirs for this opportunistic pathogen. Traditional plate culture methods remain the regulatory standard for monitoring Legionella, but these methods are slow—often requiring 7–10 days for results—and suffer from overgrowth by non-Legionella bacteria. Additionally, traditional methods fail to detect viable but non-culturable (VBNC) bacteria—cells that remain infectious but do not grow on standard culture media.
Molecular methods like PCR-based detection provide faster and more sensitive Legionella identification. However, a key limitation persists: PCR detects DNA from both live and dead bacteria, leading to false positives and unnecessary or even wasteful remediation efforts. To address this challenge, Promega has developed a viability qPCR method that retains the speed of molecular testing while distinguishing viable bacteria from non-viable remnants. In this third blog in our Legionella blog series, we cover how molecular detection methods can be refined to provide actionable results for Legionella monitoring.
Continue reading “No More Dead Ends: Improving Legionella Testing with Viability qPCR”Strengthening Water Safety Measures with Advanced Detection
Detecting Legionella in water systems is a critical step in preventing outbreaks of Legionnaires’ disease. However, not all detection methods are created equal. One of the biggest challenges in water testing is differentiating between viable and non-viable cells. This distinction is essential for making informed decisions about water system safety and compliance, especially in high-stakes environments like hospitals, office buildings and public spaces.
In a previous blog, we explored the history and significance of Legionella testing, from its discovery during the 1976 outbreak to the risks posed by modern water systems. We also highlighted the limitations of traditional culture-based detection and the need for advanced tools to improve accuracy and speed. In this second blog, we will dive deeper into the challenges of Legionella detection, the science behind qPCR technology and how an innovative approach to qPCR addresses these challenges. Finally, we will demonstrate how this technology fits into established workflows to deliver reliable, actionable results for water safety.
Understanding and Combating Legionella in Water Systems with Viability PCR
Water plays a vital role in countless aspects of daily life—drinking, cooling, recreation and more. However, the same systems that deliver these benefits can also harbor Legionella, a waterborne bacteria responsible for Legionnaires’ disease, a severe form of pneumonia (1). Legionella thrives in stagnant aquatic environments, many of which are human-made and common in modern infrastructure, like in cooling towers, hot tubs and complex building water systems. In this blog, we explore the risks posed by Legionella, the limitations of traditional detection methods and how advanced tools at Promega are transforming water safety monitoring.
Hot Off the Seep: Novel Cyanobacteria with Hefty Implications for Carbon Cycling
Cyanobacteria, microscopic photosynthetic bacteria, have been quietly shaping our planet for billions of years. Responsible for producing the oxygen we breathe, these tiny organisms play a critical role in the global carbon cycle and are now stepping into the spotlight for another reason: their potential to both understand and potentially combat climate change.
Recently, researchers discovered two new strains of cyanobacteria, UTEX 3221 and UTEX 3222, thriving in a marine volcanic seep off the coast of Italy. While cyanobacteria are virtually everywhere there is water and light—from calm freshwater ponds to extreme environments like Yellowstone’s hot springs—this particular habitat is remarkable for its naturally high CO₂ levels and acidic conditions. For these newly identified strains, a geochemical setting like marine volcanic seeps have likely driven the evolution of unique traits that could make them valuable for carbon sequestration and industrial applications.
Continue reading “Hot Off the Seep: Novel Cyanobacteria with Hefty Implications for Carbon Cycling”Why Do We Love Being Scared? The Science Behind Horror Movies
There’s something oddly captivating about watching a film that makes you jump, scream, or better yet—a film that sticks with you long after watching. Millions of people embrace the fear, willingly diving into the dark world of horror movies. But why? What is the appeal of subjecting ourselves to terror? The reasons we watch and enjoy scary movies go far beyond the jump scares—they’re deeply psychological.
For those who find themselves covering their eyes or clutching the nearest pillow, it might be hard to understand. Yet, as the hair-raising month of October ends, many people spent the 31 days leading up to Halloween watching films designed to scare the daylights out of them. In this blog, we explore why people enjoy fear (or why they don’t) and what psychology reveals about the movies that truly terrify us.
Continue reading “Why Do We Love Being Scared? The Science Behind Horror Movies”iGEM Grant Winners Tackle Tough Problems in Synthetic Biology
In June, Promega proudly announced the ten winners of the 2024 Promega iGEM Grant. These extraordinary teams have been hard at work preparing for the iGEM Grand Jamboree, which will take place from October 23-26, 2024, in Paris, France. We interviewed a handful of this year’s grant recipients to learn more about their projects and journeys they’ve taken to reach this exciting milestone. Below are stories from four of the winning teams.
