Personal
Projects
Here you will find a compilation of the research essays and noteworthy projects that I have spent my Undergraduate years as a Biology student working on in order to improve my ability to think as a scientist and gain a greater appreciation for the world of science.
During my time studying Psychology, I undertook a challenge to delve deeper into a topic that I personally enjoy and write a research article. The topic was regarding the human mind, or more specifically how memories can essentially be recreated into a false memory to in a way "hack" the mind. By researching on the long and short term memories and the ways through which those storage units within the brain could be altered, this research essay aims to demonstrate how such a complex organ as the brain could be susceptible to manipulation.
CRISPR Genetic Editing of Saccharomyces Cerevisiae
Due to my molecular biology lab course becoming remote due to COVID-19 restrictions, I was provided materials to conduct my own CRISPR experiment to edit a specific gene (ADE2) in Yeast cells from the species Saccharomyces Cerevisiae. In summary of the experiment, I constructed a research article write-up highlighting the key discoveries. The gene's natural phenotype is observed to show a white pigment, but when mutated via CRISPR, loss-of-function (LOF) mutations should yield a red color. The CRISPR mechanism solely introduces breaks within the DNA in specific predetermined spots of the gene, and the cell repairs the damage using either Nonhomologous End Joining (NHEJ) which essentially glues together the broken pieces and is significantly error prone (leading to a potentially high likelihood for LOF within cells), or Homology Directed Repair (HDR) which uses a foreign piece of DNA that when introduced into the cell's genome via natural repair mechanisms induces LOF mutations within ADE2 yielding a red color.
Above is a picture of the materials used for the experiment. On the left are the organic materials used such as culture plates, bacterial samples, and vials containing the genetic code for the CRISPR system to input within the Yeast cells. On the right contain the mechanical tools such as a pipette, inoculating loop, L-shaped cell spreader, and most importantly gloves.
For this experiment, it is vital to establish a control to ensure that the cells growing contain the necessary genetic blueprints to construct the CRISPR system. The culture plate labeled "Empty pML104" shows high amounts of normal cell growth identified by the white colored colonies on the plate. This proves that the cells have taken up the tools needed to grow and utilize the introduced CRISPR genetic "blueprints".
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Next, to ensure that the experiment was performed carefully with no chance of contamination, a second control is performed placing no DNA in the culture plate. A successful result will yield no growth at all which is what is observed in the plate labeled "No DNA."
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For the variables stage of the experiment, a culture plate was provided with the CRISPR system and HDR to place specific mutations that will yield LOF in the cells producing a red color, if successful. Based on the results of the above plate labeled "pML104 +gRNA+HDR" the cells show both white and red colors meaning that CRISPR genetic editing in this environment was fairly successful. It is important to note that in biology, nothing is ever rarely guaranteed!
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When the HDR component is removed, and the resulting DNA damages are to be repaired via NHEJ, the resulting observations depict that a higher level of red colonies appear relative to the white indicating that CRISPR editing has resulted in a greater LOF within this environment. Overall, this experiment has demonstrated a fairly successful attempt at genetically editing the ADE2 gene in Yeast cells to produce a red color as opposed to a white color during growth.
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