SERGEY B. KUZNETSOV

**** *** **. **********, ** *2980

434-***-**** ( abob72@r.postjobfree.com

Experienced Molecular and Cell Biologist

EDUCATION / CREDENTIALS

Ph.D., Genetics, Institute of Cytology & Genetics, Novosibirsk, Russia.

Master of Science, Biology, Rostov State University, Rostov-on-Don,

Russia.

(Both Ph.D. and M.S. were qualified by Global Credential Evaluators Inc.

and Morningside Evaluations & Consulting for

accreditation in USA

AREAS OF EXPERTISE

. Strong experience spanning molecular and cell biology discovery, cell

pluripotency and differentiation, cancer biology, gene construction,

and recombinant protein expression.

. Solid background in mouse and mouse embryo manipulations, in vivo/ex

vivo procedures, and development of molecular diagnostic and cell-

based functional assays for drug discovery.

. Qualified to initiate and direct laboratory operations, research, and

experiments supporting pre-clinical and clinical studies. GLP, SOP.

. Developed communication skills with talent for inspiring superior team

performance while coordinating initiatives in deadline-driven

environments.

. Author of Several Publications, in genetics and cancer research.

. Cell Line Development, Cell and Tissue Culture

. DNA & RNA Isolation, Analysis, Detection

. PCR/RT-PCR, DNA Sequencing

. Cloning & DNA Library

. Western Blotting and protein electrophoresis, ELISA

. Recombinant Protein Expression

. Mammalian & Bacterial Cells Transfection/Transformation

. Gene Engineering Laboratory Protocols & Reporting

. Mouse & Mouse Embryo Manipulation

. Mouse Xenograft Cancer Models, Knock-out Mice

. Cryosectioning of Mammalian Tissues

. Immuohistochemistry & Microscoping

. Scientific Research & Development, Scientific Writing & Editing

PROFESSIONAL EXPERIENCE

UNIVERSITY OF LOUISVILLE, Louisville, Kentucky ( 2010-2011

Research Associate, Gheens Center of Aging: Planned and executed research

projects focused on the role of micro RNAs in the processes of development

and aging. Conducted in-depth analysis and quantitative/qualitative

research of critical subject matter data and processes. I routinely

utilized

common molecular biology techniques such as PCR, restriction enzyme

cleavage, ligation, etc. to construct plasmid and viral DNA constructions,

and used them to produce stable cell lines in aceptic conditions.

Highlights:

. Isolated and cultured mouse primary neuron cells.

. Designed and built plasmid DNA constructions with different micro RNA

genes for target delivery to the mouse brain for neuron specific gene

expression.

UNIVERSITY OF RHODE ISLAND, Kingston, Rhode Island ( 2008-2010

Research Fellow, Department of Physics: Performed cutting-edge cancer

research encompassing in vivo imaging of tumors and direct delivery of

specific drugs to treat tumors. Utilized mouse xenograft

model of cancer; injected and grew human cancer cells, and dissected

tumors/organs for cryosectioning and microscoping. I used several human

tumor cell lines to develop solid tumors in nude mice via subcutaneous

injections, and IV injections of such cells to imitate metastatic process.

After developing of tumors I injected (IV and IP) solution of specially

designed peptide labeled with fluorescent dye to deliver this peptide

directly to the tumors to make these tumors visible in specific wavelength

light. The effectiveness of delivery I checked with the use of Kodak in

vivo imaging system. Then I dissected the tumors, kidneys, liver, and some

other organs to make cryosectioning, and investigate an internal

distribution of labeled peptide in these organs and tissues under

fluorescent microscope. Also, I used the peptide labeled with gold

nanoparticles to deliver them directly to the tumors, and investigated

their distribution in tumors, kidneys, and liver on cryosections of these

organs. I studied also the pharmacokinetics of injected materials in mice.

During several days after injections I checked the dynamics of releasing of

injected materials in 13 organs and tissues using in vivo, ex vivo imaging,

and microscoping.

Highlight:

. Produced findings illuminating the possibility to mark tumors and

metastases with special fluorescent dyes, enabling detection in organisms

and effective dissection of tumors during surgical operations.

UNIVERSITY OF VIRGINIA, Charlottesville, Virginia ( 2005-2008

Research Associate, Department of Pathology: Served as key contributor to

high profile research initiatives focused on the role of NORE1A gene and

protein (relative to RASSF1A gene) in tumor suppression, and the mechanisms

of this suppression. Monitored and maintained compliance with data

collection, and project protocols. Prepared and submitted comprehensive

reports, documentation, and research results. I routinely utilized DNA/RNA

and protein isolation, PCR, RT-PCR, Western blotting to construct plasmid

and viral DNA constructions, and used them to produce stable cell lines. I

used ELISA to evaluate a level of protein expression and xenograft models

of cancer to estimate an effect of tumor growth suppressing of NORE1A

protein. I designed and constructed plasmids with NORE1A gene and

established stable human tumor cell lines using Flow Cytometry and

antibiotic selection. I used immunohystochemistry techniques to estimate

the intracellular localization of NORE1A protein, and shRNAs to knockdown

the gene. Then I injected transfected cells into nude mice to check the

tumor suppressive ability of this protein. In the study I found that

several tumor cell lines can specifically degrade the over expressed NORE1A

protein, and confirmed this with RT-PCR and Western blotting. Later I found

the same effect in several tumors from patients of UVA Hospital. All of

these findings allowed us to describe the new mechanism of escaping of

growth suppression in tumor cells.

Highlights:

. Successfully confirmed the suppressive activity of NOR1A protein on mouse

model, and discovered a new tumor self-protection mechanism from NORE1A

and RASSF1A suppressors.

. Developed plasmid DNAs; transfecting human and mouse cancer cells,

checking the expression of proteins of interest.

. Delivered conclusive evidence that calpains, a family of proteins

involved in growth control, are responsible for cleavage of NORE1A and

its closest relative RASSF1A tumor suppressor by tumor cell lines.

. Identified a new mechanism allowing many lung cancer cells to escape

growth suppression: a proteolysis of tumor suppressors NORE1A and RASSF1A

by calpains as alternative mechanism of their down-regulation.

UNIVERSITY OF CONNECTICUT HEALTH CENTER, ( 2003-2005

Visiting Fellow, Center for Vascular Biology: Pivotal driver in the study

of Hif2a (hypoxia inducible factor 2, alfa subunit) gene and protein, a key

contributor to the vascularization of tumors. Analyzed a difference in

expression and degradation of this protein and its close relative Hif1a

gene and protein. I routinely utilized common molecular biology techniques

to construct plasmid and viral DNA constructions, and used them to produce

stable cell lines and knocked-out mice routinely used PCR for genotyping of

mouse embryos, and confocal microscopy to investigate defects of blood

vessels development in them. I developed an approach for visualization of

blood vessels using direct injections of fluorescent solution into heart of

7-8 days embryos. Also, I maintained the colony of knockout mice.

INSTITUTE OF CYTOLOGY & GENETICS, Novosibirsk, Russia ( 1986-2003

Senior Research Scientist (1996-2003): Played vital role in numerous large-

scale research projects. Designed experiments; developed and maintained

laboratory protocols, notebooks, and documentation. Effectively formulated

and executed strategic design, preparation, and conduct of study. Applied

for and won multiple grants from Russian Fund of Basic Investigations, as

well as an Emergency Research Grant from the International Science

Foundation. I was head of student graduation projects and co-director of

dissertation projects. I headed the research team to map the genes of

mammals and the study of pluripotency and reprogramming of differentiated

cells within 5 years. I routinely utilized different

molecular biology and biochemical techniques, such as protein

electrophoresis and hystochemical stainings, Western blotting, PRC and RT-

PCR, DNA/RNA isolation and sequencing, etc.

Highlights:

. Successfully completed cutting edge research and projects in the areas of

gene mapping, and cell and chromosome differentiation study in hybrid

somatic cell clones, and somatic hybrids of embryonic stem cells and

differentiated cells between different mammalian species, including

human.

. One of a select group of individuals assigned to Grant # 265 from Russian

Government to support young scientists: "Mapping of pig chromosomes using

pig-mink hybrid cells: chromosome 2 and 12".

. Deployed molecular and biochemical methods (PCR/RT-PCR; protein

electrophoresis; etc) for precise gene mapping in mammals, and

investigation of cell differentiation.

. Credited with developing numerous techniques to detect smallest

differences in electrophoretic mobility of enzymes of parental species in

hybrid clones, which was critical for gene mapping of the pig genome.

Subsequently, using somatic cell hybrid panel pig-American mink,

singlehandedly mapped 20 genes of the domestic pig, Sus scrofa.

. Localized more 50+ genes and microsatellite loci of the American mink,

Mustella vison, with the somatic cell hybrid panel American mink-Chinese

Hamster, in collaboration with Dr. S. O'Brien from the National Cancer

Institute, a world leader in comparative gene mapping.

. Played major role in the study of pluripotency of embryonic stem cells by

using hybrid cell panel between mouse embryonic stem cells and

differentiated cells. Delivered groundbreaking results including

discovery of non-random segregation of "differentiated cell" chromosomes

in the hybrid cells and a phenomenon of "chromosomal memory".

Note: Additional positions with Institute of Cytology & Genetics includes

Research Scientist, Research Fellow, and Research Assistant.

LIST OF PUBLICATIONS

1. Yao L, Danniels J, Moshnikova A, Kuznetsov S, Ahmed A, Engelman DM,

Reshetnyak YK, Andreev OA.. The pHLIP peptide targets nanogold

particles to tumors. Proc Natl Acad Sci U S A. 2013 Jan 8;110(2):465-

70

2. Reshetnyak Y.K. L.Yao, S. Zheng, S. Kuznetsov, D. Engelman, O.

Andreev. Measuring tumor aggressiveness and targeting metastatic

lesions with fluorescent pHLIP. Mol Imaging Biol. 2011 Dec;13(6):1146-

56.

3. Kuznetsov Sergey, Andrei Khokhlatchev. The Growth and Tumor

Suppressors NORE1A and PASSF1A Are Targets for Calpain-Mediated

Proteolysis. PLoS ONE. www.plosone.org December 2008, V. 3, Issue 12,

e3997 http://www.plosone.org/article/info%3Adoi%2F10.137

1%2Fjournal.pone.0003997

4. Moshnikova A., Kuznetsov S., Khokhlatchev A.V. "Interaction of the

growth and tumour suppressor NORE1A with microtubules is not required

for its growth-suppressive function". BMC Research Notes, 2008, 1:13

5. Larkin D.V., Astakhova N.M., Kozhin A.Yu., Kuznetsov S.B., Zhdanova

N.S. The structure of a conservative region of porcine genome,

represented in a human genome by chromosome 17. Russian J. of

Genetics. 2004, -v.40(7), P. 782-788

6. Sergei B. Kuznetsov, William J. Murphy, Stephen J. O'Brien, and Oleg

L. Serov. Mapping of 53 loci in American mink (Mustela vison). Journal

of Heredity 2003 Sep-Oct; 94(5):386-91

7. Serov O.L., Matveeva N.M., Kizilova E.A., Kuznetsov S.B., Zhelezova

A.I., Golubitsa A.N., Pristyazhnyuk I.E., Puzakov M.V. Chromosome

memory" of parental genomes in embryonic hybrid cells. Russian Journal

of Developmental Biology. 2003 May-Jun; 34(3):175-184. Russian.

8. Serov OL, Matveeva NM, Kizilova EA, Kuznetsov SB, Zhelezova AI,

Golubitsa AN, Pristiazhniuk IE, Puzakov MV. Differentiated cell

chromosomes. An.Acad.Bras.Cien. 2001, -v. 73(4), P. 561-568.,

9. Serov O, Matveeva N, Kusnetsov S, Kaftanovskaya E, Mittmann J.

Embryonic hybrid cells: a powerful tool for studying pluripotency and

reprogramming of the differentiated cell chromosomes. An Acad Bras

Cienc. 2001, 73:561-568.

10. Serov O.L., Matveeva N.M., Kuznetsov S.B., Kaftanovskaya E.M.

Embryonic stem cells: new possibilities in investigation of

pluripotency and reprogramming of chromosomes of differentiated cells.

Izvestiya Academii Nauk (Biologiya) (Russia). 2001, -v. 6, P. 711-716.

11. N.M.Matveeva, S.B.Kuznetsov, E.M.Kaftanovskaya, O.L.Serov. Segregation

of the parental chromosomes in murine hybrid cells obtained by fusion

of embryonic stem cells with splenocytes. Proceedings of Russian

Academy of Sciences (Russia) 2001, -v.379/1, P. 118-120.

12. Larkin D.M., S.B.Kuznetsov, N.M.Astakhova, N.S.Zhdanova. The using of

PCR markers for mapping of pig chromosome 12. Genetica (Russia).

2001. -37, P. 358-364.

13. Ivanova E.V., I.V. Koroleva., S.B. Kuznetsov, T.I. Axenovich, G.P.

Svisheva, S.N. Malchenko, C. Bendixen, N.S.Zhdanova.

Radiation map of short arm of pig chromosome 2. Genetica (Russia)

2001. -v.37, P.-230-237.

14. Koroleva I.V., S.N. Malchenko, N.M. Shukri, I.V. Ivanova, N.M.

Astakhova, S.B. Kuznetsov, N.S. Zhdanova, C. Bendixen. Assignment of

six comparative anchor loci to pig chromosome 2 by pig-mink cell

hybrids. Mammalian Genome, 1998, v. 9.

15. Kuznetsov S.B., D.M. Larkin, E.M. Kaftanovskaya, E.V.Ivanova, N.M.

Astakhova, O.V.Cheryaukene, N.S.Zhdanova. Chromosomal localization and

analysis of synteny of several genes in pig, cattle, and sheep

(Artiodactyla). Genetica (Russia). 1998.V.34 (9). P.1-5.

16. Rubtsov N E.V. Plyusnina, Graphodatsky A., S?rdukova N.V., N.M.

Astakhova., S.B. Kuznetsov., Zhdanova, N.S. New possibilities of

analysis of complex chromosomal rearangements in cell hybrids.

Genetica (Russia). 1998. V.34.P.240-247.

17. Koroleva I.V., Malchenko S., Shukri N.M. Kuznetsov S.B., Ivanova E.V.,

Zhdanova N.S., Bendixen C. Assignment of 5 porcine genes by pig-mink

hybrids with single pig chromosomes 2, 5, 8, 12. Mamm. Genome, 1998.

-v. 9, -P. 913-914.

18. Zhdanova, N. S., P. D. Tomsen, E. V. Plyusnina, N. M. Astakhova,

S.B. Kuznetsov, S.B. Jorgensen, and O.L.Serov. Partial collection of

pig-mink cell hybrids with single pig chromosomes. Archivos de

zootecnia, -1996. -v. 45, -P. 367-370

19. Zhdanova N. S., P.D.Thomsen, N.M. Astakhova, S.B. Kuznetsov,

E.V.Plyusnina, O.L.Serov. Production of pig-mink cell hybrids with

single pig chromosomes 2, 5, 12 or t(1.13). Mammalian Genome, 1996,

-v.7, -P. 613-615.

20. Kuznetsov S.B., Baranyuk V.V., Takekawa J.Y. Genetic differentiation

between subpopulations of Lesser Snow Geese nesting on Wrangel Island,

Russia. Auk, 1998. V. 115, No.4, -P. 1053-1057.

21. Kuznetsov S.B. Polymorphism of blood plasma esterases in the geese

of Anser genus (AVES: ANSERIFORMES). Biochem. Genet., -1995. -v. 33,

5/6. -P. 183-187.

22. Kuznetsov S.B. Polymorphism of blood plasma proteins in the geese of

Anser and Branta genera. Biochem. Genet., -1995. -v. 33, 3/4. -P. 123-

135.

23. Astakhova N.M., N.S.Zhdanova, E.M.Kaftanovskaya, S.B.Kuznetsov,

O.L.Serov. Preliminary

assignment of genes TK1 and UMPH2 to pig chromosome 12. Genetica.

1994, -v. 30, -P. 832-838. (In Russian).

24. Zhdanova N.S., N.M.Astakhova, S.B.Kuznetsov, L.Schuler, O.L.Serov.

Characterization of pig-mink cell hybrids: assignment of the TK1 and

UMPH2 genes to pig chromosome 12. Mammalian Genome, 1994. -v. 5. -P.

781-784.

25. Kuznetsov S.B. Polymorphism of blood plasma proteins in the geese of

Anser and Branta genera. Siberian Biological Journal (Russia). 1991,

-v.5, P.3-10.

PRESENTATION AND LECTURES

1. Moshnikova A., Kuznetsov S., Khokhlatchev. A. Identification of the

Aurora A kinase as an interacting partner for the growth and tumor

suppressor NORE1A". Minisymposium session "Cellular and Molecular

Biology 69" at the 2008 AACR Annual Meeting in San Diego, CA, April

16, 2008.

2. Moshnikova A., Kuznetsov S., Khokhlatchev A. Interaction of the RASSF

family growth and tumor suppressor NORE1A with microtubules and

centrosomes. Mechanisms and Models of Cancer. La Jolla, CA. August 8-

12, 2007.

3. Moshnikova A.B.,. Kuznetsov S B., Khokhlatchev A.V. Interaction of the

growth and tumor suppressor NORE1A with microtubules. AACR Annual

Meeting 2007, April 14-18, Los Angeles,CA V.48, p. 102.

4. Matveeva N.M., Kuznetsov S.B., Shilov A.G., Kaftanovskaya E.M.,

Zhelezova A.I., Golubitsa A.N., Kizilova E.A., Serov O.L. The study of

pluripotency and segregation of parent chromosomes in hybrids of

embryonic stem cells and somatic cells. Russian symposium "Biology of

stem cells", Moscow, November 15-17, 2000.

5. Ivanova E.V., Kuznetsov S.B., Koroleva I.V., Bendixen Chr., Zhdanova

N.S. A radiation hybrid map of porcine chromosome 2p. Plant and Animal

Genome VIII Conference. San Diego, January 9-12, 2000.

6. Zhdanova N.S., Ivanova E.V., Kuznetsov S.B., Axenovich T.I., Svisheva

G.R., Astakhova N.M. The mapping of pig chromosome 2. Abstracts of II

Congress of Vavilov's Genetic Society. Sankt-Petersburg, 2000. -v 2.

P. 37.

7. Zhdanova N.S., Larkin D.M., Kuznetsov S.B., Rubtsov N.B., Astakhova

N.M. The order of genes on porcine chromosome 12. 27 international

conference on animal genetics. Conference abstract book. Minneapolis,

July 22-26, 2000. P.52.

8. N.M.Matveeva, A.G. Shilov, H.M.Kaftanovskaya, L.P.Maximovsky,

A.I.Zhelezova, A.N.Golubitsa, S.B.Kuznetsov, O.L.Serov. Pluripotent

enbrionic stem-somatic cell hybrids containing a single somatic x-

chromosome. International Symposium on X-chromosome Inactivation in

Mammals. September 6-12, 1999, Novosibirsk, Russia (Poster Abstracts)

9. Larkin D.M., Kuznetsov S.B., Ivanova E.V., Astakhova N.M., Serov O.L.,

Zhdanova N.S. Comparative mapping of human chromosome 17 genes in pigs

and shrews. 11th North American Colloquium on Domestic Animal

Cytogenetics and Gene Mapping. July 14-18, 1999, University of

Minnesota. (Abstracts)

10. Kuznetsov S.B., Rubtsov N.B., Larkin D.M., Ivanova E.V., Astakhova

N.M., Zhdanova N.S. Microdissection of chromosomes in regional mapping

of unique genes. 11th North American Colloquium on Domestic Animal

Cytogenetics and Gene Mapping. July 14-18, 1999, University of

Minnesota. (Abstracts)

11. Larkin D.M., S.B. Kuznetsov, H. Kaftanovskaya, H. Ivanova, N.

Zhdanova. Mapping of several HSA17 genes in pig, cattle, and sheep.

The International Conference on the Status of Plant and Animal Genome

Research. January 18-22, 1998, Town and Country Hotel, San Diego, CA.

(Poster Abstracts)

12. Kuznetsov S.B. Population processes and relationships between two

populations of Lesser snow geese nesting on Wrangel Island.

International Conference on new methods of investigations in natural

populations. Moscow, 1995. (Oral report)

13. Zhdanova N.S., N.M. Astakhova, S.B. Kuznetsov, O.L.Serov. Porcine

region of synteny homology in human chr 17 and mouse chr 11. Abstracts

of oral and poster papers. 2-nd European Congress of mammalogy, 27

march-1 april, 1995. Southgampton, 1995. (Poster session)

14. Kuznetsov S. B. Baranyuk V. V. The genetic differences between two

subpopulations of Lesser Snow Geese nesting on Wrangel Island, Russia.

Abstracts of 8-th North American Arctic Goose Conference and

Workshop. Albuquerque, New Mexico. 1995. (Poster session)

15. Zhdanova N.S., N.M. Astakhova, S.B. Kuznetsov, O.L. Serov Assignment

of TK1 and UMPH2 genes to pig chromosome 12. XXIV International

Conference on Animal Genetics, Prague, Czech Republic, 1994, P.132.

(Poster session)

16. Kuznetsov S.B.. Polymorphism of blood plasma proteins in Anser and

Branta genera (Aves: Anseriformes). XXIV International

Conference on Animal Genetics, Prague, Czech Republic,

1994, P.41.

17. Kuznetsov S.B. The electrophoretical analysis of proteins and enzymes

of Anser and Branta geese. Abstracts of 7-th North American Arctic

Goose Conference and Workshop. Vallejo, California. 1992. (Poster

session)

18. Sharshov A.A., Kuznetsov S.B. Comparative cytogenetics of Anser geese

(Aves: Anseriformes). 3 Workshop on genetics and selection of animals.

Biysk, 1989. Izvestiya Sibirskogo otdeleniya Akademii Nauk.

Biologicheskaya Seriya (Russia). 1989, -v.2, P. 53.

Referees

Oleg Serov, DS, Professor

Head of Laboratory of Developmental Genetics at the

Institute of Cytology and Genetics (Novosibirsk, Russia)

Russian Academy of Science,

Siberian Branch

630090 Novosibirsk, RUSSIA

abob72@r.postjobfree.com

Office: +7-383-***-****

Andrei Khokhlatchev, PhD

Senior Scientist University Of Virginia

Department: Md-Cell Cell Biology

Office Phone: +1-434-***-****

E-mail: abob72@r.postjobfree.com ; abob72@r.postjobfree.com

Ludmila Matyakhina, Ph.D., FACMG

Cytogenetics, Director

GeneDx

207 Perry Parkway

Gaithersburg, MD 20877

Phone: 301-***-**** ext 6183

E-mail: abob72@r.postjobfree.com

Contact this candidate