From strawberry DNA extraction to gel electrophoresis and everything in between, I have learned so much and have had an amazing time here at Fred Hutch this summer. Ever since I was young, I have been drawn to science, especially biology and the human body. So when I got the opportunity to participate in this program, I knew this would be a summer to look forward to.
Me in our lab measuring reagents with
a micropipette.
Two of the labs that stood out to me were the PCR of the B2M gene and the CRISPR-cas 9 digest. The B2M gene is a part of the body’s Major Histocompatibility Complex or the “self-identifying” section of the immune system. In relation to cancer detection, this gene identifies cancer cells as a part of the body and allows them to grow and spread. In order to look into this gene more, we needed to make more copies of it which is where the PCR comes in. PCR, or Polymerase Chain Reaction, is a technique used to make many copies of a particular section of DNA. The steps of PCR are Denaturation, Annealing, and Extension. Denaturation is where the strands of DNA are separated. Annealing is where Primers are attached to the DNA. This prepares for Extension, which is where Taq polymerase builds on the attached primers to create a new strand of DNA. After copying this gene and looking further into it, we decided that in order for the body to detect cancer cells and trigger a proper immune response for everyone, this gene needed to be cut. For that we needed to use CRISPR-cas 9. This is a new gene editing tool that can edit genetically programmed immune system cells called CAR T-cells to edit the B2M gene so that it triggers a universal immune response to cancer within the CAR T-cell. Through this, these programmed immune system cells can be more affordable and accessible to patients (specifically patients with cancerous B cells). During this gene editing process, the cas 9 protein and a single strand of a guide RNA work together to look for a genetic sequence called the PAM sequence in the base pairs of the DNA. Once they find this sequence the CRISPR unzips the DNA on the opposite side of the PAM sequence to see if it is complementary to the guide RNA. Once the sequence is proved to be complementary, the guide RNA binds with the DNA of the opposite side and the cas 9 protein makes a cut in the DNA 3 base pairs in from the PAM sequence. Finally, the proteins around the DNA looking from problems in the genes begin any needed gene repair.
Results of our PCR, the microtubule furthest Results of our CRISPR digest in Agarose Gel.
The right shows the DNA glowing.
These labs specifically stood out to me because they showed me just how much we can do with the technology we have in medical labs and how big of an impact it makes in the treatment of patients. With the process of PCR, we have a gateway into discovery that we did not have before. Through the ability to multiply genes, we are able to catch any new pieces of information about this gene that can be a crucial starting point to a treatment. By finding these starting points we are able to advance more quickly in providing treatments. With the CRISPR cas-9 gene editing tool, we can manipulate our body to work for us in a way that were not possible before. Both of these advancements are helping people live their happiest and healthiest lives. The fact that work being done in the lab is having a positive impact on the lives of people is incredible!
On a final note, I would like to thank Dr. Kay Lalish, Dr. Gennifer Goode, Hanako, Jesse,Tayla, Dr. Raabya Rossenkhan, my peers who participated in this program with me, and everyone else who was a part of making this program possible. It was truly a memorable experience!
Me and my friend Faduma waiting for result Me and my friends Melody and Faduma
in the lab waiting for results in the lab