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MicroRNAs and Prostate Cancer

MicroRNAs and Prostate Cancer. By: Carol Tran Dr. Zendra Zehner Dr. Xueping Zhang. MicroRNA Review. http://www.jyi.org/articleimages/392/originals/img2.jpg. Project Goals.

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MicroRNAs and Prostate Cancer

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  1. MicroRNAs and Prostate Cancer By: Carol Tran Dr. Zendra Zehner Dr. Xueping Zhang

  2. MicroRNA Review http://www.jyi.org/articleimages/392/originals/img2.jpg

  3. Project Goals To test if miR23 and/or miR17 have an effect on the growth, tumorigenic, or metastatic properties of prostate cancer cells.

  4. Prostate Gland Cross-Section F6 M12

  5. miR23 on Chromosome 19

  6. Summer Goal 1.) To monitor the levels of the microRNAs in M12 compared to the F6 subline. 2.) To change the levels of the microRNA and measure the differences in the reporter gene activity. http://www.clontech.com/products/detail.asp?product_id=10606&tabno=2

  7. 5’ 3’

  8. Process • Clone DNA • Transform Bacterial Cells to take up Plasmid DNA molecule • Mini-Preps and Maxi-Preps • Monitor Effect on Reporter Gene Activity- Transfection • Assays for Results

  9. miR Sequence hsa-miR-23a: ATCACATTGCCAGGGATTTCC 5’GATCCGATCACATTGCCAGGGATTTCCTTTTCAAGAGAGGAAATCCCTGGCAATGTGATTTCTTTTTTTCTAGAG gCTAGTGTAAGGTCCCTAAAGGAAAAGTTCTCTCCTTTAGGGACGTTACACTAAAGAAAAAAAGATCTCTTAA 5’ hsa-miR-23a-AS: GGAAATCCCTGGCAATGTGAT 5’GATCCGGGAAATCCCTGGCAATGTGATTTTTCAAGAGAATCACATTGCCAGGGATTTCCTTCTTTTTTTCTAGAG gCCCTTTAGGGACCGTTACACTAAAAAGTTCTCTTTAGTGTAACGGTCCCTAAAGGAAGAAAAAAAGATCTCTTAA 5’

  10. miR17

  11. mir23

  12. Future Ideas • Could measure exact amount of miRs in M12 and F6 sublines instead of reporter gene activity. • Northern Blot and radioactivity

  13. CONCLUSION • Poured plates • Mini plasmid preps • Maxi plasmid preps • Took ODs • dual-Luciferase Assay • Culture Cells • Transfections

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