Poster #037, University of California, Riverside
Engineering U87 Cells to Modulate Stress Resilience through Extremophile Protective Proteins
Mentors: Joshua Morgan, PI; Sara Nafar, Mentor; Daniel Bustamante, Mentor
Tardigrades and brine shrimp can survive extreme conditions, including radiation, temperature extremes, and complete dehydration. They achieve this through cryptobiosis, a reversible state of suspended metabolism. This tolerance is partly attributed to the expression of stress proteins unique to each organism such as: CAHS3 and MAHS in tardigrades, and AfrLEA3m in brine shrimp. CAHS3 stabilizes cells under stress by forming a reversible gel-like network. MAHS preserves mitochondrial function during stress. AfrLEA3m prevents aggregation of key mitochondrial enzymes during dehydration.
In this project, we expressed these three proteins in the U87 cell line, a human astrocyte model, to establish a platform for future evaluation of their stress-protective potential in a mammalian system. Gene delivery was performed using the Sleeping Beauty transposon system, a non-viral method that uses SB transposase to stably integrate genes into the host genome. The primary goal was to generate stable U87 lines expressing CAHS3, MAHS, or AfrLEA3m, alongside a control line, as a foundation for subsequent stress-tolerance studies in human cells.