This summer we’re sponsoring high school interns in stem cell labs throughout California. We asked those students to contribute to our Instagram photos and YouTube videos about life in the lab, and write about their experiences.
Alfonso Barraza did a stem cell research internship this summer in the laboratory of Jan Nolta at the University of California, Davis.
|Alfonso Barraza working in the lab. He submitted this photo to our #CIRMStemCellLab Instagram feed.|
First of all hello. My name is Alfonso Barraza. I have been working in the Nolta Lab at the UC Davis Institute for Regenerative Medicine. I have pretty much my whole summer been working with the one and only Vector Goddess, also known as Karen Pepper. First day was alright (not really). I mean I got to meet Scary G which was pretty awesome. To be honest I was kind of scared to meet him (don’t tell him that) because he is the UC Davis program director. I really didn’t know what to expect. But I tell you once you get to know him, he is awesome. I also had to go through all of the paperwork and “necessary” stuff for working there. There was a lot less than I thought. THANK GOODNESS!! Going through biosafety training was enough work in itself. Don’t get me wrong the lady who was teaching us everything and telling us what to do when you spill chemicals was really nice. But it’s really hard to make something like biosafety training fun and enjoyable (I almost fell asleep).
The next day was awesome because I finally got to meet her Vectorness. Boy is she awesome. She knows everything about vectors and I had the privilege of learning about vectors from her. There was just so much information; lentiviruses, retroviruses, promoters, suicide genes, backbones, and what all of the letters and numbers mean on her eppendorf tubes.
I finally learned what my project was the next day, 4XBDNF as I like to call it. What if I want to produce higher amounts of protein? What are the options? Well higher MOI’s (more virus per cell) could be used, but the FDA won’t allow that because it could cause problems like insertional mutagenesis and cancer. So other options? Why don’t I just add additional copies of a gene to a single virus? So that’s what I did. Because Brained Derived Neurotrophic Factor (BDNF) is a highly characterized protein in lab due to its importance in Huntington’s Disease research, this concept was based around the BDNF gene. I did all of my DNA work with the Goddess herself. I must of tried at least 7 times to get that stubborn cassette of 4 BDNF genes to clone into the vector backbone. It just wouldn’t go in. Finally it went in and I was so grateful because I finally got to start packaging my vector.
I began working with Catherine Nacey to package my vector, because a virus doesn’t work if its just a plasmid. I got culture cells, transfect cells, and eventually harvest my virus. This probably was the best part because I had created my very own vector.
I then needed to try it to see if it worked, right? So I tranduced some Mesenchymal Stem Cells with my virus to see how much BDNF it produced. AND drum roll please…… The cells transduced with my virus didn’t produce anymore BDNF than the wild-type cells (CRIES)!!!! Yep. It didn’t work and me, my mentor, and Dr. Fernando Fierro all have no idea why it failed. It kinda sucks it didn’t work, but what can you do.
I just want to say thanks to all of the awesome people in the Nolta Lab, Dr. Jan Nolta, Dr. Gerhard Bauer, Catherine Nacey, Kari Pollock, Dr. Fernando Fierro, and the Goddess for everything.
(P.S. I guess I’ll have to come back next summer to fix it!)
Alfonso submitted this video about his experience: