SPARK Research…on Gene Therapy Vectors! by Lauren Duan

It’s been a crazy couple months. We started where every budding scientist starts, with little knowledge but lots of curiosity. Now that we’re near the end of our time in the CIRM SPARK program, it feels incredibly rewarding to be able to see that we can walk out of our respective labs knowing a little bit more about the world of regenerative medicine and a lot more about what research truly entails.
My experience during this internship was a bit different from others: my project didn’t even involve true stem cells, but rather focused on gene therapy vectors. Whereas my peers could talk about their MSC, hESC, or iPSC cultures, I could only nod and be glad we learned enough about these stem cells in our graduate-level Bio225 class to hold a conversation. I work with HEK293T cells that come from a line of transformed cells that originated from human embryonic kidney cells. They’re mostly used as producer cells for things such as viral vectors, which can be used for gene therapy to insert new functionaDuan, Lauren IMG_3170l genes to cure currently incurable diseases!

The main focus of my project was to find a more efficacious way to purify viruses, specifically adeno-associated virus, from all of the other cell debris. The two methods I tested are purification through Zeba desalting columns and fast protein liquid chromatography (FPLC). At the end, we found out which one works better by “counting” the relative amounts of DNA in the same volume of sample using qPCR, which should correlate in a 1:1 ratio with the amount of virus, since each virus has one single strand of DNA. The method with the highest amount of DNA would then be the most effective at purification! By finding the best way to purify the virus, I’ll be able to contribute to creating a more effective method of delivering virus-mediated gene therapy.

Duan, Lauren IMG_4022 cropIf I said I work with highly concentrated viral vector suspensions everyday while wearing a full body suit, two pairs of gloves taped to my sleeves, and a face mask, it might seem like I’m doing something extremely dangerous. However, if all goes well and the gene therapy vector works as it should, I really would only be at risk of being cured of a disease! Working in the Good Manufacturing Practice (GMP) facility at the UC Davis Institute for Regenerative Cures means that personal protective equipment (PPE) really does protect both me and the product.

The GMP is like no other place I’ve ever been. With fewer than 10,000 particles larger than 0.45 microns per cubic foot per minute in each manufacturing room (while the outside air has around 35,000,000 on the same scale), I’ve never been in a place so clean. Working with the GMP employees, being able to see and participate in clinical-grade manufacturing, has been absolutely invaluable. It’s really allowed me to connect all of the experimental science done in the basic and translational research labs to the actual therapies that go into patients during clinical trials.

Duan, Lauren IMG_4065 crop

I can’t say enough how grateful I am for this opportunity to see research from the eyes of a real scientist. Even though I’m young and liberated from most responsibility and commitment, I can safely say that my future in science is as sure as can be.

 

 

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