Health Molecular Biology

Min-Hyuk Yoo, Ph. D

**** ******** **** ****

Rockville, MD, 20852

301-***-**** (home) or 703-***-**** (cell)

acp1wo@r.postjobfree.com

Permanent resident

CORE EXPERTISE

Over 15 years' experience and knowledge of biological science

Expertise and creativity in cell and molecular biology techniques

- mammalian cell culture, cell transfection, and cell-based functional assays,

- Expertise in designing and executing immunological methods such as ELISA, FACS

and immunostaining

- Experience with molecular biology such as cloning, mutagenesis and PCR

Strong cell and molecular biology experience such as tissue culture techniques, transfection, RNAi knockdown (siRNA), protein and gene reporter assays, RNA/DNA/protein extraction, and western blotting for translational research projects

Excellent written and verbal communication, work planning, data analysis and record keeping skills

Ability to independently design experiments, make detailed observations, analyze data, interpret results and compile summary reports

Enthusiastic team player with cooperative and responsible personality, good organizational and communication skills with other research teams

SUMMARY OF ACCOMPLISHMENTS

Awarded US patent (US 2010/0034835 A1, 2010, " Use of inhibitors of leukotreiene B4 receptor BLT2 for treating asthma") for treating Asthma

Proved the involvement of thioredoxin reductase 1 (TR1) in cancer characteristics : tumor formation, metastasis, self sufficient growth, resistance to extracellular stress

Demonstrated the involvement of Glutathione Peroxidase 4 (GPx4) with the onset of Alzheimer's disease

Participated in the development of stable multiple knockdown strategy using RNAi knockdown (siRNA) technology for characterizing function of gene(s)

Optimized various laboratory experimental procedures for accuracy, efficiency and reproducibility of data : cloning, immunoblot, immnoprecipitation, metabolic labeling with isotopes, flow cytometry (FACS), and immunofluorescence microscopy

PROFESSIONAL EXPERIENCE

National Cancer Institute, Laboratory of Cancer Prevention - Bethesda, MD

Staff Scientist September 2010 - April 2014

Elucidated the role and the mechanism of thioredoxin reductase 1 in increased resistance of cancer cells against extracellular stresses using knockdown cells by siRNA

Proved the role of TR1 in specific selenite toxicity to cancer cells

Collaborated with other scientists to examine the role of TR1 in various cellular mechanism

Wrote manuscripts for publication in scientific journals

National Cancer Institute, Laboratory of Cancer Prevention - Bethesda, MD

Visiting Fellow September 2004 – September 2010

Elucidated the role of TR1 in various cancer phenotypes such as anchorage independent growth, self-sufficient growth signal, metastasis and tumor formation

Discovered the functional role of glutathione peroxidase 4 (GPx4) in oxidative stress induced-membrane damage repair

Supervised postdoctoral staff in project initiation, progression and publication

Wrote manuscripts for publication in scientific journals

Set up and optimized liver and breast cancer mouse model

Kwangju Institute of Science & Technology, Department of Life Science

- 123 Chumdangwagi-ro, Buk-gu, Gwangju, 500-712, Korea (www.gist.ac.kr)

Doctoral Research Fellow September 1999 - February-2004

Proved the role of leukotriene B4 and its receptor (BLT2) in cancer progression

Proposed the regulation of leukotriene B4 pathway as therapeutic strategy of asthma

Set up Ovalbumin (OVA) induced asthma mouse model : harvest and analysis of bronchoalveolar lavage fluid (BAL) fluid, airway hyperresponsiveness (AHR) and bronchial biopsy specimen, preparing mouse intraperitoneal macrophage and bone marrow cells

EDUCATION

2004 Ph.D., Life Science, Department of Life Science, Kwangju Institute of Science & Technology, Korea (Thesis: Study on the Role of Leukotriene-linked Cascade in Human Disease)

1999 M.S., Biochemistry, Graduate School of Biotechnology, Korea University, Korea (Thesis: Effects of Selenium on Growth of Human Lung Cell Lines)

1997 B.S., Genetic engineering, Department of Genetic engineering, Korea University, Korea

TECHNICAL SKILLS

1. Molecular biology

More than 10 years of gene cloning and mutagenesis for over-expression or reporter gene assay

Stable RNA interference (RNAi) vector construction for single or multiple genes

RT-PCR, northern blot, microarray, immunoprecipitation, immunofluorescence microscopy

Electrophoretic mobility shift assay (EMSA) for measuring activity of NF-kB or HIF-1a

2. Biochemistry

Ten years of western blot, luciferase assay

Thioredoxin resuductase, glutathion peroxidase activity assay with mouse tissue or mammalian cells

3. Cell biology

Fifteen years of experience with various mouse and human cell lines and primary cells

Transient and stable transfection for overexpression or knockdown of specific gene or genes.

Chemotaxis assay with various cells, reporter gene assay, cell cycle analysis

Fluorescence-activated cell sorting (FACS) for analysis of membrane protein, intracellular ROS and cell cycle

4. Mouse disease model

General skills such as subcutaneous (SC), intraperitoneal (IP) injection, oral gavage, harvest various tissues.

Prepare and culture mouse peritoneal macrophages and bone marrow cells.

Maintain transgenic and tissue specific knockout mouse: genotyping.

Tumor formation assay with cultured mammalian cancer cells.

Ovalbumin (OVA) induced asthma mouse model: harvest and analysis of bronchoalveolar lavage fluid (BAL) fluid, airway hyperresponsiveness (AHR) and bronchial biopsy specimen, preparing mouse intraperitoneal macrophage and bone marrow cells.

DMBA (7,12-dimethylbenz(a)anthracene) induced breast cancer model: breast cancer cell line establishment from tumor tissues.

DEN (diethylnitrosamine) induced liver cancer mouse model.

LANGUAGES

Fluent in English and Korean

PATENTS

Kim JH, Cho KJ, Yoo MH, Use of inhibitors of leukotreiene B4 receptor BLT2 for treating asthma. (US 2010/0034835 A1, 2010)

Invited Presentations

1. Yoo MH, Carlson BA, Tsuji PA, Gladyshev VN, and Hatfield DL: Role of thioredoxin reductase 1 in oxidative stress in cancer cells, FASEB Experimental Biology, San Diego CA, 2012.

2. Yoo MH, Carlson BA, Gladyshev VN, and Hatfield DL: Targeted Removal of Thioredoxin Reductase 1 Increases TNF-a induced Apoptosis in Breast Cancer Cells, 9th International Symposium on Selenium in Biology and Medicine. Kyoto, Japan, 2010.

3. Yoo, MH, Xu, XM, Carlson, BA, Gladyshev, VN, and Hatfield, DL. Increased sensitivity of thioredoxin reductase 1 deficient breast cancer cells against TNF-a induced apoptosis, FASEB Experimental Biology, New Orleans LA, 2009.

4. Yoo MH, Patterson AD, Xu XM, Carlson BA, Gladyshev VN, and Hatfield DL: Functional analysis of phospholipid hydroperoxide glutathione peroxidase by targeting its expression, FASEB Experimental Biology, San Diego CA, 2008.

5. Yoo MH, Xu XM, Carlson BA, Gladyshev VN, and Hatfield DL: Examination of anticancer mechanisms in ras-induced cancer cells by targeting thioredoxin reductase 1 knockdown, FASEB Experimental Biology, Washington DC, 2007.

6. Yoo MH, Xu XM, Turanov AA, Carlson BA, Gladyshev VN, and Hatfield DL: Thioredoxin Reductase 1 is a Molecular Target for Cancer Therapy and a Knockdown/Knock-in Model for Studying Selenoprotein Function, 8th International Symposium on Selenium in Biology and Medicine. Madison, WI, 2006.

7. Yoo MH, Xu XM, Turanov AA, Carlson BA, Gladyshev VN, and Hatfield DL: siRNA knockdown-mRNA knock-in as a means of assessing selenoprotein function, FASEB Experimental Biology, San Francisco, CA, 2006.

Chapters in Books

1. Carlson BA, Yoo MH, Tsuji PA, Tobe R, Naranjo-Suarez S, Chen F, Feigenbaum L, Tessarollo L, Lee BJ, Gladyshev VN, and Hatfield DL. Mouse models that target removal or over-expression of the selenocysteine tRNA gene to elucidate the role of selenoproteins in health and development. In: Selenium: Its molecular biology and role in human health, (3rd Edition), Hatfield DL, Berry MJ, Gladyshev VN (Eds.), Chap 44, 3rd Edition. Springer Science+Business Media, LLC, New York, 2011.

2. Yoo MH, Carlson BA, Tsuji PA, Tobe R, Naranjo-Suarez S, Lee BJ, Davis CD, Gladyshev VN, and Hatfield DL. Selenoproteins harboring a split personality in both preventing and promoting cancer. In: Selenium: Its molecular biology and role in human health (3rd Edition), Hatfield DL, Berry MJ, Gladyshev VN (Eds.), Chap 25, 3rd Edition. Springer Science+Business Media, LLC, New York, 2011.

3. Xu XM, Turanov AA, Carlson BA, Yoo MH, Gladyshev VN, and Hatfield DL. Selenocysteine biosynthesis and the replacement of selenocysteine with cysteine in the pathway. In: Selenium: Its molecular biology and role in human health (3rd Edition), Hatfield DL, Berry MJ, Gladyshev VN (Eds.), Chap 2, 3rd Edition. Springer Science+Business Media, LLC, New York, 2011.

4. Carlson BA, Xu XM, Shrimali R, Sengupta A, Yoo MH, Zhong N, Hatfield DL, Irons R, Davis CD, Lee BJ, Novoselov SV and Gladyshev VN : Mouse models for assessing the role of selenoproteins in health and development. In: Selenium--Its Molecular Biology and Role in Human Health (2nd Edition), Hatfield DL, Berry MJ, and Gladyshev VN (Eds.) Chap 29, 2nd Edition, Springer+Business Media, LLC, New York, NY, 2006.

5. Carlson BA, Xu XM, Shrimali R, Sengupta A, Yoo MH, Irons R, Zhong N, Hatfield DL, Lee BJ, Nobanov AV and Gladyshev VN: Mammalian and other eukaryotic selenocysteine tRNAs. In: Selenium--Its Molecular Biology and Role in Human Health (2nd Edition), Hatfield DL, Berry MJ and Gladyshev VN (Eds.), Chap 3, 2nd Edition, Springer+Business Media, LLC, New York, NY, 2006.

Publications

1. Yoo MH, Carlson BA, Gladyshev VN, Hatfield DL. Abrogated thioredoxin system causes increased sensitivity to TNF-a-induced apoptosis via enrichment of p-ERK 1/2 in the nucleus. PLoS One 8(9): e71427, 2013.

2. Naranjo-Suarez S, Carlson BA, Tobe R, Yoo MH, Tsuji PA, Gladyshev VN, Hatfield DL. Regulation of HIF-1α activity by overexpression of thioredoxin is independent of thioredoxin reductase status. Mol. Cells 36(2):151-157, 2013.

3. Patterson AD, Carlson BA, Li F, Bonzo JA, Yoo MH, Krausz KW, Conrad M, Chen C, Gonzalez FJ, Hatfield DL. Disruption of Thioredoxin Reductase 1 Protects Mice from Acute Acetaminophen-Induced Hepatotoxicity through Enhanced NRF2 Activity. Chem. Res. Toxicol. [Epub ahead of print], 2013.

4. Tobe R, Naranjo-Suarez S, Everley RA, Carlson BA, Turanov AA, Tsuji PA, Yoo MH, Gygi SP, Gladyshev VN, Hatfield DL. High error rates in selenocysteine insertion in mammalian cells treated with the antibiotic, doxycycline, chloramphenicol or geneticin. J. Biol. Chem. 288(21):147**-*****, 2013.

5. Tsuji PA, Carlson BA, Naranjo-Suarez S, Yoo MH, Xu XM, Fomenko DE, Gladyshev VN, Hatfield DL, Davis CD. Knockout of the 15 kDa Selenoprotein Protects against Chemically-Induced Aberrant Crypt Formation in Mice. PLoS One 7(12): e50574, 2012.

6. Carlson BA, Yoo MH, Tobe R, Mueller C, Naranjo-Suarez S, Hoffmann VJ, Gladyshev VN, and Hatfield DL: Thioredoxin reductase 1 protects against chemically induced hepatocarcinogenesis via control of cellular redox homeostasis. Carcinogenesis 33(9): 1806-1813, 2012.

7. *Tobe R, *Yoo MH, Fradejas N, Carlson BA, Calvo S, Gladyshev VN, and Hatfield DL: Thioredoxin reductase 1 deficiency enhances selenite toxicity in cancer cells via a thioredoxin-independent mechanism. Biochem J. 445(3): 423-430, 2012. *1st co-author

8. Naranjo-Suarez S, Carlson BA, Tsuji PA, Yoo MH, Gladyshev VN, and Hatfield DL: HIF-independent regulation of thioredoxin reductase 1 contributes to the high levels of reactive oxygen species induced by hypoxia. PLoS One 7(2): e30470, 2012.

9. Turanov AA, Xu XM, Carlson BA, Yoo MH, Gladyshev VN, and Hatfield DL: Biosynthesis of selenocysteine, the 21st amino acid in the genetic code, and a novel pathway for cysteine biosynthesis. Adv. Nutr. 2(2):122-128, 2011.

10. Tsuji PA, Naranjo-Suarez S, Carlson BA, Tobe R, Yoo MH and Davis CD: Deficiency in the 15 kDa selenoprotein inhibits human colon cancer cell growth. Nutrients 3(9): 805-817, 2011.

11. Carlson BA, Yoo MH, Conrad M, Gladyshev VN, Hatfield DL, and Park JM: Protein kinase-regulated expression and immune function of thioredoxin reductase 1 in mouse macrophages. Mol. Immunol. 49 (1-2): 311-316, 2011.

12. Xu XM, Turanov AA, Carlson BA, Yoo MH, Everley RA, Nandakumar R, Sorokina I, Gygi SP, Gladyshev VN, and Hatfield DL: Targeted insertion of cysteine by decoding UGA codons with mammalian selenocysteine machinery. Proc. Natl. Acad. Sci. U S A. 107(50): 214**-*****, 2010.

13. Carlson BA, Yoo MH, Shrimali RK, Irons R, Gladyshev VN, Hatfield DL, and Park JM: Role of Selenium-containing proteins in T-cell and macrophage function. Proc. Nutr. Soc. 69(3): 300-310, 2010.

14. Turanov AA, Kehr S, Marino SM, Yoo MH, Carlson BA, Hatfield DL, and Gladyshev VN: Mammalian thioredoxin reductase 1: roles in redox homoeostasis and characterization of cellular targets. Biochem. J. 430(2): 285-293, 2010.

15. Irons R, Tsuji PA, Carlson BA, Ouyang P, Yoo MH, Xu XM, Hatfield DL, Gladyshev VN, Davis CD: Deficiency in the 15 kDa Selenoprotein Inhibits Tumorigenicity and Metastasis of Colon Cancer Cells. Cancer Prev. Res. 3(5): 630-639, 2010.

16. Yoo MH, Carlson BA, Xu XM, Tsuji PA, Irons R, Gladyshev VN, and Hatfield DL: Alteration of Thioredoxin Reductase 1 Levels in Elucidating Cancer Etiology. Methods Enzymol. 474: 255-275, 2010.

17. Yoo MH, Gu X, Xu XM, Kim JY, Carlson BA, Patterson AD, Cai H, Gladyshev VN, and Hatfield DL: Delineating the role of glutathione peroxidase 4 in protecting cells against lipid hydroperoxide damage and in Alzheimer’s Disease. Antioxid. Redox. Signal. 12(7): 819-827, 2010.

18. Cho KJ, Seo JM, Shin Y, Yoo MH, Park CS, Lee SH, Chang YS, Cho SH and Kim JH: Blockade of airway inflammation and hyperresponsiveness by inhibition of BLT2, a low-affinity leukotriene B4 receptor. Am. J. Respir. Cell Mol. Biol. 42(3): 294-303, 2010.

19. Carlson BA, Yoo MH, Tsuji PA, Gladyshev VN, and Hatfield DL: Mouse models targeting selenocysteine tRNA expression for elucidating the role of selenoproteins in health and development. Molecules 14(9): 3509-3527, 2009.

20. Carlson BA, Yoo MH, Sano Y, Sengupta A, Kim JY, Irons R, Gladyshev VN, Hatfield DL, and Park JM: Selenoproteins regulate macrophage invasiveness and extracellular matrix-related gene expression. BMC Immunol. 10(1): 57, 2009.

21. Xu XM, Yoo MH, Carlson BA, Gladyshev VN, and Hatfield DL: Simultaneous knockdown of the expression of two genes using multiple shRNAs and subsequent knock-in of their expression. Nat. Protoc. 4(9):1338-1348, 2009.

22. Labunskyy VM, Yoo MH, Hatfield DL, and Gladyshev VN: Sep15, a Thioredoxin-like Selenoprotein, is Involved in the Unfolded Protein Response and Differentially Regulated by Adaptive and Acute ER Stresses. Biochemistry 48(35): 8458-8465, 2009.

23. Hatfield DL, Yoo MH, Carlson BA, and Gladyshev VN: Selenoproteins that function in cancer prevention and promotion. Biochim. Biophys. Acta. 1790(11):1541-1545, 2009.

24. Yoo MH and Hatfield DL: The Cancer Stem Cell Theory: Is It Correct? Mol. Cells 26(5):514-516, 2008.

25. Yoo MH, Xu XM, Turanov AA, Carlson BA, Gladyshev VN, and Hatfield DL: A new strategy for assessing selenoprotein function: siRNA knockdown/knock-in targeting the 3'-UTR. RNA 13(6):921-929, 2007.

26. Yoo MH, Xu XM, Carlson BA, Patterson AD, Gladyshev VN, and Hatfield DL: Targeting thioredoxin reductase 1 reduction in cancer cells inhibits self-sufficient growth and DNA replication. PLoS One 2(10):e1112, 2007.

27. Yoo MH, Xu XM, Carlson BA, Gladyshev VN, and Hatfield DL: Thioredoxin reductase 1 deficiency reverses tumor phenotype and tumorigenicity of lung carcinoma cells. J. Biol. Chem. 281(19): 130**-*****, 2006.

28. Yoo MH, Song H, Woo CH, Kim H, and Kim JH: Role of BLT2, a leukotriene B4 receptor, in Ras Transformation. Oncogene 23: 9259-9268, 2004.

29. Woo CH, Yoo MH, You HJ, Cho SH, Mun YC, Seong CM and Kim JH: Transepithelial migration of neutrophils in response to leukotriene B4 is mediated by a reactive oxygen species-extracellular signal regulated kinase-linked cascade. J. Immunol. 170: 6273-6279, 2003.

30. Lee MN, You HJ, Cho SH, Woo CH, Yoo MH, Joe EH, and Kim JH: Implication of the small GTPase Rac1 in the generation of reactive oxygen species in response to beta-amyloid in C6 astroglioma cells. Biochem. J. 366: 937-943, 2002.

31. *Jeong DW, *Yoo MH, Kim TS, Kim JH, and Kim IY: Protection of mice from allergen-induced asthma by selenite: prevention of eosinophil infiltration by inhibition of NF- B activation. J. Biol. Chem. 277: 178**-*****, 2002. *1st co-author

32. Eom YW, Yoo MH, Woo CH, Hwang KC, Song WK, Yoo YJ, Chun JS, and Kim JH: Implication of the small GTPase Rac1 in the apoptosis induced UV in Rat-2 fibroblasts. Biochem. Biophys. Res. Commun. 285: 825-829, 2001.

33. Yoo MH, Woo CH, You HJ, Cho SH, Kim BC Choi JE, Chun JS, Jhun BH, Kim TS and Kim JH: Role of the cytosolic phospholipase A2-linked cascade in signaling by an oncogenic, constitutively active Ha-Ras isoform. J. Biol. Chem. 276: 246**-*****, 2001.

34. Woo CH, Eom YW, Yoo MH, You HJ, Han HJ, Song WK, Yoo YJ, Chun JS and Kim JH: Tumor necrosis factor-alpha generates reactive oxygen species via cytosolic phospholipase A2-linked cascade. J. Biol. Chem. 275: 323**-*****, 2000.

35. Woo CH, Kim BC, Kim KW, Yoo MH, Eom YW, Choi EJ, Na DS and Kim JH: Role of cytosolic phospholipase A2 as a downstream mediator of Rac in the signaling pathway to JNK stimulation. Biochem. Biophys. Res. Commun. 268: 231-236, 2000.

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