C University

Yi Hsing Lin

Irvine, CA, *****

Cell: 206-***-****

*********@*****.***

Summary:

• Great working knowledge and hands on skills in protein characterization (western blot, SDS

page, HPLC…etc.), and molecular biology.

• Extensive experiences in protein purification including IMAC, IEX, and SEC.

• Proficient in lentivirus production, titration, and infection.

Professional experiences:

Research Scientist III Dept. of Pharmacology, University of Washington 2007-2012

• Purified five His tagged antibodies expressed in E.coli and validated the purified antibodies by

ELISA and flow cytometry in two months.

• Overexpressed protein in HEK 293 and CHO cultured cells for enzymatic activity assays.

• Validated and analyzed high throughput screening data within 2 months to find the targets.

Constructed and overexpressed HA and GFP tagged proteins to validate the targets.

• Designed, optimized, and performed qPCR to detect and quantify gene expression in cultured

cells/mouse brain

• Knocked down protein expression by lentivirus shRNA in cultured/primary cells. Transferred

the knocked down cells for enzymatic studies.

• Designed and optimized high throughput siRNA screening to identify small molecules targeting

GPCRs for the treatment of Glioblastoma Multiforme.

Senior fellow

Division of Medical Genetics, University of Washington 2006-2007

● Constructed, overexpressed and purified the recombinant GST tagged protein followed by conducting

DNA-protein binding studies in three months.

Postgraduate Researcher

Dept. of Medicine, University of California at San Diego 2003-2004

● Quantified PAI I protein expression by immunoblotting and ELISA.

Postdoctoral Fellow

Dept. of Immunology, The Scripps Research Institute, La Jolla, California 2000-2003

● Identified proteins enriched in purified pseudopodia by MudPIT (multidimensional protein

identification technology ) .

● Performed cell-based assays/Immunoassays /molecular biology/immunocytochemistry to

elucidate protein binding partners/signaling pathways.

Education

Ph.D. Dept. of Biochemistry, McGill University, Montreal, Canada.

● Purified to homogeneity from bacteria crude extract a new protein after five major chromatography

steps, affinity chromatography step and reverse-phase HPLC.

M.Sc Department of Agriculture Chemistry, National Taiwan University, Taipei, Taiwan

• Purified recombinant Aspartase in E.coli followed by enzyme kinetics assays.

B.Sc. Department of Agriculture Chemistry, National Taiwan University, Taipei, Taiwan

Technical Skills:

Molecular biology: RT-PCR, PCR, Real time PCR (Stratagene MX3000 and Light CyclerT),

multiplex PCR, Qantitative Gene expression assays, Microarray analysis, Gene silencing using

lentivirus and siRNA. High throughput siRNA functional screening, DNA sequencing, molecular

cloning, Chromatin immunoprecipitation., library constructions, Southern and northern hybridization,

mobility shift assays, reporter gene assays, library constructions, site-directed mutagenesis, yeast two

hybrids screening.

Protein Purification/Biohemistry: Protein expression, western blot, protein engineering, FPLC

(IEX, SEC), HPLC, Affinity columns, SDS-PAGE, Protein stabilizations, enzyme kinetics, radioligand

binding assays, subcellular fractionation.

Cell Biology: mammalian cell cultures, Flow cytometry, transient transfection, and development of

stable cell lines, Western blot, ELISA, immunoprecipitation, cell migration assays and wound healing

assays. Apoptosis assays. Toxicity assays. Kinase assay, Immunocytochemistry/immunohistochemistry,

confocal/deconvolution microscopy,.

Bioinformatics: Strong expertise with commercial molecular biology software such as Vector NTI and

Primer Design …etc.. Extensive experiences working with bioinformatics database such as BLAST

(sequence alignment), NCBI, GenBank, genome.ucsc…etc.. .

Microbiology: various Bacteria and yeast culture, drug resistance assays, Gene replacement.

Addendum

Honors and Awards:

• California Breast Cancer Research Program postdoctoral fellowship: Lasp-1 Signaling in

Breast Carcinoma Cell Invasion/Migration. 2001-2003. Award: $85,508

• C.F. Wang McGill Major Fellowship: 1993-1995.

Scientific Publications:

1. Downregulation of cannabinoid receptor 1 from neuropeptide Y interneurons in the basal ganglia of

patients with Huntington's disease and mouse models. Horne EA, Coy J, Swinney K, Fung S, Cherry

A.E., Marrs W.R., Naydenov A.V., Lin Y.H., Sun X, Dirk Keene C, Grouzmann E, Muchowski P,

Bates G.P., Mackie K, Stella N. Eur J Neurosci. (2012 Nov 21.) doi: 10.1111/ejn.12045. [Epub ahead

of print]

2. Dual inhibition of alpha/beta-hydrolase domain 6 and fatty acid amide hydrolase increases

endocannabinoid levels in neurons. Marrs WR, Horne EA, Ortega-Gutierrez S, Cisneros JA, Xu C, Lin

Y.H., Muccioli GG, Lopez-Rodriguez ML, Stella N. J Biol Chem. (2011) 286(33):28723-8.

3. NIR-mbc94, a fluorescent ligand that binds to endogenous CB(2) receptors and is

amenable to high-throughput screening. Sexton M, Woodruff G, Horne EA, Lin Y.H.,

Muccioli GG, Bai M, Stern E, Bornhop DJ, Stella N. Chem Biol. (2011) 18(5):563-8.

4. ABHD6 controls 2-AG accumulation and efficacy at cannabinoid receptors. Marrs WR,

Blankman JL, Horne EA, Thomazeau A, Lin Y.H., Coy J, Bodor A, Muccioli GG, Hu SS,

Woodruff G, Fung S, Lafourcade M, Alexander JP, Long JZ, Li W, Xu C, Mackie K, Manzoni

OJ, Cravatt BF and Stella N .(2010) Nat. Neurosci. 13(8):951-7.

5. Binding of NIR-conPK and NIR-6T to astrocytomas and microglial cells: evidence for a

protein related to TSPO. (2009) Sexton M, Woodruff G, Cudaback E, Kreitzer FR, Xu C, Lin Y.H.,

Möller T, Bai M, Manning HC, Bornhop D, Stella N. PLoS One. 18;4(12):e8271.

6. Differential changes in GPR55 during microglial cell activation. Pietr M, Kozela E, Levy R,

Rimmerman N, Lin Y.H., Stella N, Vogel Z, Juknat A.(2009) FEBS Lett. 583(12):2071-6.

7. Lin Y.H., Park Z.-Y., Brahmbhatt A., Rio, M.-R., Yates, J., and Klemke R.L. (2004) Regulation of

cell migration and survival by focal adhesion targeting of Lasp-1. J. Cell Biol. 165: 421-32. Faculty

1000. Score 3.0.

8. Lin Y.H., Miyamoto, C., and Meighen, E.A. (2002) Cloning, sequencing, and functional

studies of the rpoS gene from Vibrio Harveyi. Biochem. Biophy.Res. Commun. 293:456-62.

9. Lin Y.H., Miyamoto, C., and Meighen, E.A. (2000) Cloning, gene replacement and

phenotype characterisation of luxT, a transcriptional factor of luxO from V. harveyi.

Biochim. Biophys. Acta. 1494: 226-35.

10. Lin, Y.H., Miyamoto, C., and Meighen, E.A. (2000) Purification and characterization of a

luxO promoter binding protein LuxT from V. harveyi. Protein Expr. Purif. 20: 87-94.

11. Miyamoto,C.M., Lin,Y.H., Meighen, E.A. (2000) Control of Bioluminescence in Vibrio fischeri by

the LuxO Signal Response Regulator. Mol Microbiol, 36:594-607.

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