Examining gene expression with RT-qPCR

At Visikol, our services have grown from our expertise in cell culture, high-content tissue imaging, and advanced digital pathology.  For nearly a year, however, our in vitro services have been augmented by gene expression analyses that allow us to provide clients with a more complete overview of the often complex interactions lurking behind the results produced by drug candidates.  These assays center around quantitative (real-time) reverse transcription polymerase chain reaction, thankfully known day to day as RT-qPCR.

The polymerase chain reaction’s common nickname of “PCR” has worked its way into conversation for many around the world during the course of the global coronavirus pandemic.  PCR’s use in COVID-19 testing has granted it a recognizable place in public medical awareness alongside mainstay three-letter abbreviations like MRI, ACL, and UTI.  However, while it may come up at the dinner table a lot more often these days, PCR in various forms has been at the core of molecular biology for decades and truly revolutionized the field after its invention by Kary Mullis in 1983.  (Mullis would go on to win the Nobel Prize in Chemistry a decade later, and the story of his PCR “revelation” has become a part of modern scientific folklore.  Seriously, look it up sometime…!)

The concepts that form the foundation of PCR are deceptively simple.  With minimal resources – basically a few simple reagents and a way to control their temperature – researchers are able to take any sequence of DNA and recreate it billions of times, essentially generating as many copies of a sequence of interest as they desire.  The process is fast, consistent, relatively inexpensive…and undeniably beautiful.  Indeed, Dr. Mullis was like Lennon and McCartney in a lab coat, reaching into thin air and pulling out something magical that had always existed but was just waiting to be put in order.

Variations on the core process of PCR have led to a long list of modern applications for the platform.  At Visikol, we make extensive use of RT-qPCR to provide clients with a full picture of the effects of their test compounds.  This expansion of PCR’s capabilities adds the use of a reverse transcriptase (the “RT” in the title) to generate a DNA template from RNA samples, and its quantitative/real-time aspect (the “q”) involves step-by-step monitoring of PCR products to determine of the amount of target DNA that began the process.  Coupled with the capture of messenger RNA at critical points during the action of clients’ compounds of interest, RT-qPCR gives us a clearer picture of a drug’s impact by showing us the changes in gene expression elicited by its addition to cells.

If your company is looking for a way to solidify your understanding of a drug’s effects by complementing traditional histological and imaging techniques – or even for a first-line assessment of a compound’s impact on your tissue of interest – RT-qPCR may be the technique you’re looking for.  Our scientists can work with your requirements and budget to determine what experimental design and RT-qPCR protocol will work best for your needs.  Whether it’s for cancer drug efficacy, liver disease pathology, or a model with less well-defined gene expression parameters, we can help elaborate on the molecular phenomena lying beneath the complex mechanisms of your drug candidates.

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