1. Gene Expression Alex, Ann, Michael, Rhishi, Tish, Yaldez, Vangelis Facilitator: Jim

2. Audience This unit is intended for a mid-level, biology major’s course

3. Outline of the course - Transcription and translation - Common techniques for measuring gene expression - Introduction to epigenetics (including today’s tidbit) - Epigenetic regulation

4. Learning goal The aim of this unit is to help students understand the mechanisms and the role of epigenetics in organism development and health.

5. Learning objectives: The students will be able to: 1. Model histone – DNA interactions 2. Compare and contrast the impact of acetylation and methylation on gene expression 3. Evaluate an experiment that demonstrates epigenetic silencing 4. Discuss consequences of epigenetics on human health

6. Introduction to epigenetics

11. Electron micrograph of chromatin © 2003 Nature Publishing Group Olins, D. E. & Olins, A. L. Chromatin history: our view from the bridge. Nature Reviews Molecular Cell Biology 4, 811 (2003). All rights reserved. A B

12. Electron micrograph of chromatin © 2003 Nature Publishing Group Olins, D. E. & Olins, A. L. Chromatin history: our view from the bridge. Nature Reviews Molecular Cell Biology 4, 811 (2003). All rights reserved. A B What are the molecules at A: a) DNA b) histone c) histone, DNA and histone tail d) all of the above What are the molecules at B: a) DNA b) histone c) histone, DNA and histone tail d) all of the above

13. How can we figure out if epigenetic marks affect gene expression?

14. -> Isolate and fragment chromatin -> Antibody to grab complexes -> Design primers that flank region of interest -> Detection of amplified DNA product Chromatin immunoprecipitation (ChIP) is used to assess DNA-protein interactions

15. How can we figure out if epigenetics affects gene expression?

16. Relative enrichment of DNA GATA4 Promoter AB You perform a ChIP experiment in which you are comparing results between two tissues; brain and heart. Your PCR results are as follows: Which column do you predict represents the results from heart tissue? Remember: The amount of amplified DNA is measured and is proportional to the strength of DNA- histone interaction.

17. Relative enrichment of DNA GATA4 Promoter

18. ONOFF Alternate scenario In order to study the changes in the promoter of GATA4, scientists repeated the ChIP experiment following the treatment with the drug. Which of the following graphs describes the outcome of the experiment?

19. Relative enrichment of DNA GATA4 Promoter AB CD Which of the following graphs describes the outcome of the ChIP experiment before and after drug treatment?

20. Learning objectives: The students will be able to: 1. Model histone – DNA interactions 2. Compare and contrast the impact of acetylation and methylation on gene expression 3. Evaluate an experiment that demonstrates epigenetic silencing 4. Discuss consequences of epigenetics on human health

21. Case study for HW

22. Summative Assessment (See separate doc on wiki)