My Internship
My Internship Summary
For my LHSI internship, I interned at the Absalon Lab where I prepared biological tools to assist in the study of Plasmodium Falciparum, which is the parasite that causes malaria. My work consisted of cloning recombinant DNA that allowed for the study of specific genes in the parasite. I worked on preparing these biological tools for the study of a gene that is essential for the formation of the parasite’s nuclear envelope. In addition to cloning this recombinant DNA, I worked on a secondary project of recording and editing video lab protocols to assist with training new lab personnel. This video protocol library consists of videos demonstrating how to clone DNA, and has already been used to train new rotational students in the lab. Throughout this internship, I developed skills in medical research and solidified my plans on attending medical school to become a physician scientist.
The Absalon Lab
About the Absalon Lab
Located in the Pharmacolgy & Toxicology Departement at the IU School of medicine, the Absalon Lab is interested in studying Plasmodium Falciparum, which is the parasite that causes the most severe form of malaria. These parasites infect red blood cells, which leads to the development of many health issues. Led by Dr. Sabrina Absalon, the lab researches the nature of the asexual stage of the parasite in infected red blood cells.
The Purpose of Gene Knockdown for Understanding P. Falciparum Biology
Scientists are interested in knocking out a gene to determine if that gene is essential to the life of the organism being studied. I learned about this concept with this fun analogy of an alien visiting Earth for the first time. An alien came to earth and discovered people. The alien was fascinated about people and wanted to understand more about human physiology. After observing people for a few days, the alien observed that people used their legs to walk around and live. Wanted to find out if legs were essential for human life, it rather grotesquely removed someone's legs and observed what happened. Thankfully, the person survived, but their life was very different. The alien then concluded that legs are important for human life, but they are not necessarily essential. However, if the alien removed the heart, this would be a different story.
Much like an alien discovering a new world, scientists study the world of microbiology to determine what genes are essential for the life of disease-causing parasites. This is done by preventing the expression of specific genes in a genetic knockout study, and observing if the organism dies as a result. If the gene is essential, knock down of the gene is then used to determine the gene's function. This can lead to the development of drugs that target these essential genes and kill the parasite.
My responsibilities in the lab were to prepare the DNA tools to knock down these genes in Plasmodium Falciparum.
My Project
My role in this process of genetic knockdown was to construct the gRNA that would prevent the production of the protein the target gene transcribes. I did this by cloning specially designed DNA that would eventually be transfected into the genome of the parasite and knock down the gene of interest. I worked on preparing these biological tools for the knockdown study of a gene that is essential for the formation of the parasite’s nuclear envelope.
Enzymatic Digestion of DNA
In the DNA cloning process, two sections of DNA are combined to form new DNA. Using two restriction enzymes, I cut DNA into two fragments of desired DNA that would then be fused together with an enzyme called DNA ligase. The process of extracting the desired sections of cut DNA is done by running an agarose gel, which I describe in my Making and Running an Agarose Gel video below.
Bacterial Transformations
After ligation, the DNA is transformed into a bacteria. Each dot is a bacterial colony that contains the genetically engineered DNA. The bacteria mass produce this DNA, since it provides resistance to the antibiotic on the plate. At this point the genetically engineered DNA is harvested from a bacterial colony and is integrated into the genome of the parasite using CRISPR-Cas 9 technology. The process of transforming DNA into bacteria is demonstrated in my Bacterial Transformation video below.
My Cloning Protocol Videos
Why I Chose this Lab
I specifically chose this internship site because of its active lab research setting. The ability to learn valuable molecular biology research techniques attracted me to the Absalon lab. During my internship, I was able to learn procedures for DNA electrophoresis, PCR, and bacteria culture growths, which form the backbone of molecular biology research. Developing skills in this field was especially important to me because they bring me closer to my goal of attending medical school to become a physician scientist.
What I've Learned
Working in the Absalon Lab has exposed me to the real-world application of medical research. I've also had the privilege of working with excellent graduate students and postdocs, who have mentored me through my experiments. This experience will help me make an informed decision on attending graduate school and my future career goals.
As an LHSI intern, I have also been immersed in the scientific community and scientific 'lingo'. The frequent exposure to scientific terminology has improved my scientific communication skill, as well as my self-confidence in the lab.
This internship has also taught transferable skills for my lab courses. For example, when the lesson on buffers came up in my chemistry class I was prepared because I had been working with buffered solutions at the Absalon Lab prior to the lesson. I have seen the benefit from this internship experience in my current lab classes, and I expect my lab skills to be more developed for my future labs as well.
My Impact
Preparing the biological tools needed to study of our gene of interest has had an impact on others in the lab. My work allows for the grad students, postdoc, and my PI to have faster access to biological materials for their experiments. When my work is done successfully, weeks of preparation time can be saved for these lab personnel, allowing them to utilize their time more efficiently.
In an additional project for the lab, I recorded and edited videos for DNA cloning protocols. These videos can be seen bellow, and have been used to assist in training new rotational students in the Absalon lab.
As an intern, I have been able to help other members of my lab in many ways. In the lab, it is essential that LB media is prepared before certain experiments can be conducted. One of my responsibilities was to ensure that the lab had an adequate supply of this media each week.
Additionally, I would autoclave the biohazard waste each week. Autoclaving is a process of treating waste at high temperatures and pressure to kill any hazardous biological waste. Since the lab works with blood infected with malaria-causing parasites, proper disposal of this bio waste is essential for the safety of those who may come into contact with the waste.