Making DNA relevant and exciting in the high school classroom By Anna Heyer and Rachel Zenuk

1. Making DNA relevant and exciting in the high school classroom By Anna Heyer and Rachel Zenuk

2. What is BioME? • Is it like a biome? 2

3. Bio and Me? 3 3

4. Biology from molecules to evolution? 4 4

5. What is NSF’s GK-12 initiative? • ~100 5-year training grants nationwide • Funds grad student partnerships with K-12 public school teachers • Goal: fellows acquire communication and teaching skills, while enriching STEM education in K-12 classrooms • Training a new generation of scientists • Strengthening partnerships between universities & local school districts

6. What is the focus of BioME? • Focus: teaching the life sciences via • hands-on experiences, using a • framework of evolution & • biodiversity • We are entering the fourth of five years

7. What’s the goal of a teacher-fellow partnership? • Partners work together to develop and use classroom • teaching materials in the life sciences • Partnerships are year-long, with the fellow in the • classroom every week • The fellow is a resource with scientific expertise • The fellow isnot an aide, sub, or student teacher

8. BioME taps into fellows’ individual skills and interests. Aletris Neilis: dissecting a puma (!) at Hermosa Montessori Matt Herron: teaching molecular techniques at Tucson High Adriana Racolta: planting a garden at Liberty Elementary

9. Sweet Water Wetland Project3rd grade class at White Elementary 9

10. Liberty Elementary Garden Club 10

11. 2nd Grade Insect Projects 11

12. Summer Institute • Getting acquainted with BioME philosophy & aims • Getting partnerships underway via team time • Introducing teaching resources • Discussing traits of successful partnerships

13. BioME forges strong bonds and lasting partnerships.

14. Who runs BioME? Principal Investigators Judie Bronstein Barry Roth Stacey Forsyth Graduate Coordinator Kathleen Walker K-12 Coordinator Mary Bouley Educational Evaluator Melissa Page Melvin Hall Administration

15. Our Partnership 15 15

16. The Manduca Project 16 16

17. AP Science Field Trip • Goal- to increase rigor and interest in AP science and chemistry 17 17

18. San Pedro River • Students collected insect larva, turtles, and other aquatic specimens 18 18

19. Bob cats, pumas, and snakes...Oh My! 19

20. A great trip, but most of the sleeping happened on the bus 20 20

21. DNA Extraction 21

22. DNA Extraction • Purpose • to excite students and “hook” them in on the first or second day of school • to teach basic procedures and classroom management • to introduce the molecule we will be working with all year 22 22

23. Materials • Small vials • rubbing alcohol or ethanol 70% or higher • toothpicks • liquid dish soap in water (1:2 soap to water) • 1% salt solution 23 23

24. Procedures • Swish 15 ml of salt water for 1 minute. • Pour spit mixture into conical soap tube • Rock back and forth for 2 minutes. • Add 5-10mL of chilled ethanol to the tube. • PULL OUT THE DNA! 24

25. Summarize the procedures in your own words…. • Swish with salt water • Mix with soap • Add EtOH • Twirl with collection stick • Preserve DNA 25

26. How could use DNA extraction in your classroom? • Extensions: create DNA necklaces • Questions??? 26 26

27. Microarray Lab

28. Today’s BIG Idea • Over the last 30 years many defects in genes have been linked to cancer, each promising to be the magic in understanding and curing cancer. • We now know cancer is a multistep process, and accumulation of mutations, or genetic aberrations, allows a cell to progress to tumor and malignancy.

29. Cancer is caused by genetic mutations C A A G C T A A C T DNA Normal gene C A A G C G A A C T Single base change C A A G G C G C T A A C T Additions C T C A A G A A C T Deletions

30. Cancer is caused by genetic mutations Normal cell division Cell Suicide or Apoptosis Cell damage—no repair Cancer cell division First mutation Second mutation Third mutation Fourth orlater mutation Uncontrolled growth

31. Cancer involves MULTIPLE mutations Benign tumor cells grow only locally and cannot spread by invasion or metastasis Malignant cells invade neighboring tissues, enter blood vessels, and metastasize to different sites Time Mutation inactivates suppressor gene Cells proliferate Mutations inactivate DNA repair genes Proto-oncogenes mutate to oncogenes More mutations, more genetic instability, metastatic disease

32. Oncogenes Normal cell Normal genes regulate cell growth Oncogenes accelerate cell growth and division Cancer cell Mutated/damaged oncogene

33. Tumor Suppressor Genes Normal genes prevent cancer Normal cell Remove or inactivate tumor suppressor genes Cancer cell Damage to both genes leads to cancer Mutated/inactivated tumor suppressor genes

34. DNA Repair Genes Normal DNA repair T C G A C Base pair mismatch No cancer T C T A C A G C T G T C T A C T C T A C A G C T G Cancer A G T G A G A T G No DNA repair

35. MANY Genes are Implicated in Cancer! • Every cancer can be attributed to a different set of genetic aberrations, and different genes are either expressed or not expressed. • More than 100 different types of cancer can be found within specific organs!

36. This makes cancer treatment tricky… • Each caner has a different potential of being treated by current therapies. • For example, it has been shown cancer cells that lack p53 do not respond well to radiation therapy, and other non-malignant cells lacking p53 will progress to malignancy in response to radiation. • Thus the treatment itself can cause more cancers! p53 protein NORMAL cell Excessive DNA damage Cell suicide (Apoptosis)

37. Discussion Questions • How do you determine the function of a gene? • Which genetic aberrations have been implicated in cancer? • What cellular functions are affected (turned ON or OFF) in cancer cells, and how might these affect normal cell development?

38. What is the best way to treat cancer? • Figure out which genes are mutated and which genes are expressed or not expressed in the tissue. • Gene expression of numerous genes can be looked at by a new technique called microarray analysis.

39. What is microarray analysis?

40. Microarray analysis uses cDNA to look at gene expression DNA Transcription mRNA Reverse Transcription Translation protein cDNA 40

41. cDNA is labeled with fluorescent dyes

42. Microarray analysis shows us which genes are expressed in cancer

43. Materials • Microarray slide -- this slide contains genes involved in cancer • Disposable pipette • cDNA mixture solution -- these cDNAs were made from normal breast tissue (attached to blue dye) and breast cancer tissues (attached to red dye). • Wash solution • Color developing reagent

44. Microarray Slide

45. Procedure • Place the slide onto the paper towel. • Add enough of the cDNA solution to the slide to completely cover it, but not spill off of the slide. • Let the cDNA hybridize with the microarray slide for 5 minutes. • After the 5 minute incubation of the microarray slide with cDNA, rinse off the excess cDNA with the microarray wash solution (in squeeze bottle). • Add color solution, again enough to cover the slide but not spill over the slide. This solution is toxic so take care to not get it on you, and wash off of skin immediately. Let the color solution set for 30 sec, then wash off excess with microarray wash solution. • Record you data.

46. Results

47. Discussion • Microarray analysis shows us which genes are expresses in normal cells vs. cancer cells. • Why are some genes expressed in normal cells? • Why are some genes expressed in cancer cells? • Why are some genes expressed in both?

48. Discussion • Genes expressed in normal cells only are likely tumor suppressors or DNA repair genes. • Genes expressed in cancer cells only are likely oncogenes. • Genes expressed in both are present in both conditions.

49. THANK YOU! Questions? Contact the BIOTECH Project: Dr. Nadja Anderson nadja@bio5.org 49

50. 50 50