Cycloastragenol in Microgravity: Its Effects on Telomerase Activation and Telomere Longevity in Saccharomyces cerevisiae (the “Instant” Strain of Baker’s Yeast)

Team Members Co-Principal Investigators: Sarah Higdon, Sharmayne Siu, Lauren Takeyama, and Kyra Yamamoto Collaborators: Misako Benso, Katherine Cheng, Monica De Jesu, Gillian Ferguson, Charmayne Floyd, and Avishan Nikoui-Smith

Background Telomerase is an enzyme that lengthens telomeres and prevents the gradual degradation of coding genetic material in chromosomes.

Objective To test the effects of the Cycloastragenol drug on telomerase in Saccharomyces cerevisiae (the “instant” strain of baker’s yeast) and to qualify whether or not the telomerase works more rapidly in microgravity.

Materials SAF-Instant Instant Yeast Cycloastragenol Agar medium Phosphate buffer saline Agarose

Initial Experiments Fluorescent In Situ Hybridization (FISH) Expensive/resource consuming Difficulties including: ▫Cell wall lysis ▫Inconsistencies in yeast telomere sequences ▫Telomere primer

Experiments Preliminary Experiment #1: Whether yeast growth will be inhibited under microgravity conditions (simulated using sea plaque agarose) Preliminary Experiment #2: Whether Cycloastragenol promotes yeast growth

Preliminary Experiment #1 1.Autoclave 250 mL at 157◦ F(70◦ C) 2.Cool down to 95◦ F (35◦ C), add 1.44 mg of yeast 3.Place in LAB-LINE ORBIT Environ-Shaker at 30◦F 4.Heat up to melt agarose, pour into 500ml centrifuge tubes 5.Centrifuge for 10 min at 1500 RPM, removed excess medium/agarose 6.Tenfold serial dilution

Preliminary Experiment #2 1.Add two capsules of Cycloastragenol to 250 mL of medium and 1.44 mg of yeast, shake 2.Place in LAB-LINE ORBIT Environ-Shaker for 1.5 hour, pour into 500mL centrifuge tubes 3.Centrifuge for 10 min at 2000 RPM 4.Remove excess medium until only 100mL remaining 5.Filled four test tubes with 45mL of buffer 6.Tenfold serial dilution 7.Spectrometer

Conclusion 1.Yeast can grow in simulated microgravity conditions 2.Cycloastragenol does not negatively affect the growth of yeast or elicit abnormal responses in organisms with cell walls

Acknowledgements Mr. Wyeth Collo Dr. Steven Smith and Dr. Laura Crocitto from City of Hope Mr. Keith Derrick San Marino High School San Marino Unified School District San Marino PTSA Stephen and Mary Birch Foundation CASIS NanoRacks National Center for Earth and Space Science Education Subaru of America, Inc Mr. Fadl Saadi and Mr. Dan Torelli from the California Institute of Technology

Thank you!