Paul Wassel, lll

Mentor: Dr. Jorg Bungert

College of Medicine

"In high school, I participated in a science research internship along with a class. Since then, my passion for science has brought me to research and that continued at UF."

Major

Microbiology and Cell Science

Minor

N/A

Research Interests

  • Molecular Genetics
  • Biochemistry

Academic Awards

  • University Scholars Program (2017-2018)

Organizations

  • Student Conduct Committee
  • Men's Basketball Rowdy Reptiles

Volunteer

  • Alachua County Science Fair

Hobbies and Interests

  • Sports
  • Poker
  • Video Games

Research Description

The Effect of Integrator Knockdown on Expression of the Beta-Globin Gene Locus
My project specifically focuses on the effect of INT11 knockdown on beta-globin gene expression while also looking at the length of enhancer RNAs in the HS2 region of the LCR. We reduce expression of the Integrator complex by targeting the INT11 encoding RNA by developing mouse erythroleukemia (MEL) cell clones expressing doxycycline-inducible short hairpin RNAs (shRNAs). shRNA mediated knockdown is a procedure in which small hairpin RNAs (shRNAs) are incorporated into the cell using a viral vector, causing a knockdown in the expression of a target gene. From the beginning, shRNAs are designed to fit the target sequence of a specific mRNA. Then, the shRNA is cloned into a vector which is used to transform bacterial cells, allowing for cells only with the vector to be grown and expressed using antibiotic selectivity encoded in the vector. The next step is to make viral particles from the plasmid DNA amplified in bacteria. For this the plasmid is used to transfect a virus producer cell line. The resulting viruses are then used to infect MEL cells with these viral particles. Then, finally, MEL cells are selected taking advantage of the puromycin resistance gene encoded by the virus DNA. Two different shRNA constructs targeting different regions of the INT11 mRNA will be generated and tested. After this process, successful INT11 knockdown will be examined. Two assays that will be completed are a western blot and reverse transcription followed by qPCR. After knock-down has been confirmed I will analyze if reduced integrator expression results in elongated eRNAs associated with HS2 and in expression of the adult beta-globin gene. Chromatin Immunoprecipitation (ChIP) experiments will be conducted to further delve into the different interactions INT11 has within the globin gene locus.