State University Research

J. Kyle Krady, Ph.D.

*** ******* **. ***** ***, PA 17552 C: 717-***-**** *******@*****.***

SUMMARY

Molecular biologist with expertise in recombinant protein design and expression, RNA/DNA

isolation and manipulation and cell culture based bioassay development. Adept at the expression

of recombinant proteins from many systems, including prokaryote, mammalian and baculovirus

hosts. Looking to apply and augment these skills in a stable biotech or pharmaceutical research

position.

HIGHLIGHTS

Molecular cloning recombinant protein expression

RNA/DNA manipulation protein purification and analysis

Cell culture bioassay development

Supervisory experience oral and written communication skills

EXPERIENCE

Senior Research Scientist, Synaptic Research Inc., Baltimore MD 2010-2013

• Led a group of scientists responsible for the design, construction, expression and

purification of recombinant proteins important to the company’s research goals.

• Optimized expression conditions for a number of recombinant proteins and developed

SOPs for their scaled up purification.

• Streamlined the design of recombinant proteins allowing for better expression, saving the

company money in synthesis and purification costs.

• Developed protocols to test the biologic efficacy of expressed proteins to determine if

they were worthwhile for the company to pursue.

Assistant Professor, Pennsylvania State University, Hershey, PA 1995-2009

• Maintained and ran a basic research lab that used molecular techniques to determine the

role of inflammation and microglial activation in the etiology of neurodegenerative

diseases.

• Duties included: supervision of technicians, and mentoring of post-doctoral and graduate

students; individual and collaborative efforts to obtain funding for research; and

presentation of results at meetings of peers and in peer-reviewed journals.

• Was an early proponent of the role of inflammation and microglial activation in the

etiology of diabetic retinopathy. Research led to the use of the drug minocycline as a

putative therapeutic for diabetic retinopathy.

• In collaboration with a biotech company designed in vitro bioassays utilizing qrt-PCR

and multiplex beads to determine the efficacy of their drugs as putative treatments for

diabetic retinopathy.

• Collaborated on studies demonstrating that microglial activation is delayed in diabetic

animals subjected to hypoxic/ischemia. Research suggested a possible reason for the

poorer prognosis of diabetics to H/I events.

• Led project that used MRI as a novel non-invasive method to study the important role of

microglial activation and IL-1 signaling in the pathology of stroke. Allowed for better

data analysis using a fewer number of animals.

• Collaborated on studies designed to understand the role of microglia in Parkinson’s

disease and multiple sclerosis.

• Used differential display and PCR with degenerate primers to identify Tyrosine Kinase

proteins that were differentially activated as a result of microglial activation. Research

led to important biomarkers for stages of microglial activation.

Postdoctoral Fellow, Yale University, New Haven, CT 1990-1995

• Utilized DNAse footprint analysis, gel mobility shift assays and reporter constructs to

analyze the promoter region of the parvoviral NS-1 gene.

• Designed and performed binding assays to isolate proteins that interacted with the

NS-1 gene.

• Developed assays designed to determine proteins that interacted with the NS-1

protein.

Ph.D. Student, Pennsylvania State University, Hershey, PA 1985-1990

• Advisor: Dr. Mel Billingsley, Dissertation: Molecular Neurotoxicology of

Trimethyltin

• Developed an innovative subtractive hybridization technique to isolate mRNAs

specific to neurons damaged by the neurotoxicant TMT.

• Analyzed the resulting mRNAs using in situ hybridization, sequence analysis and

Northern blot analysis.

• Research led to the isolation and characterization of a novel protein termed Stannin.

EDUCATION

Ph.D. Pharmacology, Pennsylvania State University, Hershey, PA. 1990

B.S. Biochemistry, Pennsylvania State University, University Park, PA. 1983

PUBLICATIONS

1. Krady, J.K. Oyler, G.A., Balaban, C.D., and Billingsley, M.L. Use of avidin-biotin

subtractive hybridization to characterize mRNA common to neurons destroyed by the

selective neurotoxicant trimethyltin. Mol. Brain Res. 7:287-297, 1990.

2. Toggas, S.M., Krady, J.K., and Billingsley, M.L. Molecular neurotoxicology of trimethytin:

identification of stannin, a novel protein expressed in trimethyltin sensitive cells. Mol.

Pharmacol. 42:44-56, 1993.

3. Toggas, S.M., Krady, J.K., Thompson, T.A., and Billingsley, M.L. Molecular mechanisms of

selective neurotoxicants: studies on organotin compounds. Ann N.Y. Acad Sci 679:157-177,

1993.

4. Krady, J.K., and Ward, D.C. Transcriptional activation by the parvoviral non-structural

protein NS-1 is mediated via a direct interaction with Sp1. Mol. Cell Biol. 15:524-533, 1995.

5. Dejneka, N.S., Patenow, CM., Polavarapu, R., Toggas, S.M., Krady, J.K., and Billingsley,

M.L. Localization and characterization of stannin: relationship to cellular sensitivity to

organotin compounds. Neurochem. Int. 31:801-8115, 1997.

6. S. O’Donnell, SJ Vannucci, T Frederick, JK Krady, and TL Wood. IGF-I and

Microglia/Macrophage Proliferation in the Ischemic Mouse Brain. GLIA. 39 (1):85-97,

2002.

7. JK Krady,,A. Basu, SW Levison and RJ Milner. Differential Expression of Protein Tyrosine

Kinase Genes during Microglial Activation. GLIA. 40(1):11-24, 2002.

8. A Basu, J. Kyle Krady, M O'Malley, SD. Styren, MB Glaccum, and ST DeKosky and SW

Levison. The type 1 interleukin-1 receptor is essential for the efficient activation of microglia

and the induction of multiple proinflammatory mediators in response to brain injury. J

Neurosci. 22(14):6071-82, 2002.

9. A Basu, JK Krady, JR Enterline, and SW Levison. Transforming growth factor beta 1

prevents IL-1beta induced microglial activation, whereas TNFalpha- and IL-6-stimulated

activation are not antagonized. GLIA 40(1)109-20, 2002.

10. J Kuhlow, JK Krady A. Basu and SW Levison. Astrocytic Ceruloplasmin Expression, which

is Induced by IL-1β and by Traumatic Brain Injury, Increases in the Absence of the IL-1 Type

I Receptor. GLIA 44:76-84, 2002.

11. Price M, Lang MG, Frank AT, Goetting-Minesky MP, Patel SP, Silviera ML, Krady JK,

Milner RJ, Ewing AG, Day JR. Seven cDNAs enriched following hippocampal lesion:

possible roles in neuronal responses to injury. Brain Res Mol Brain Res 117:58-67, 2003.

12. Basu A, Krady JK, Levison SW. Interleukin-1: a master regulator of neuroinflammation. J.

Neurosci Res 78:151-156, 2004.

13. Zhang, L., Nair, A., Krady, J.K., Corpe, C., Bonneau, R. H., Simpson, I. A., and Vannucci, S.

J. Estrogen stimulates microglia and brain recovery from hypoxia- ischemia in

normoglycemic but not diabetic Female mice. J. Clinical Investigation 113: 85-95, 2004.

14. Anirban Basu*, Jelena Lazovic*, J Kyle Krady, David T Mauger, Raymond P Rothstein,

Michael B Smith and Steven W Levison. Interleukin-1 and the interleukin-1 type 1 receptor

are essential for the progressive neurodegeneration that ensues subsequent to a mild

hypoxic/ischemic injury. J. Cerebral Blood Flow and Metabolism 25: 17-29, 2005.

15. J. Kyle Krady, Anirban Basu, Colleen M. Allen, Yuping Xu, Kathryn F. LaNoue, Thomas W.

Gardner, and Steven W. Levison. (2005) Minocycline Reduces Proinflammatory Cytokine

Expression, Microglial Activation, and Caspase-3 Activation in a Rodent Model of Diabetic

Retinopathy. Diabetes. 2005 May;54(5):1559-65.

16. Alistair J. Barber, David A. Antonetti, Timothy S. Kern, Chad E. N. Reiter, Rohit S. Soans, J.

Kyle Krady, Steven W. Levison, Thomas W. Gardner, and Sarah K. Bronson. (2005) The

Ins2Akita mouse as a model of early retinal complications in diabetes. IOVS. 2005

June;46(6):2210-18.

17. Lazovic J., Basu A., Lin HW, Rothstein RP, Krady JK, Smith MB, Levison SW.

Neuroinflammation and both cytotoxic and vasogenic edema are reduced in interleukin-1

type 1 receptor-deficient mice conferring neuroprotection. Stroke. 2005 Oct;36(10):2226-31

18. Kumari R, Willing LB, Krady JK, Vannucci SJ, Simpson IA. Impaired wound healing after

cerebral hypoxia-ischemia in the diabetic mouse. J Cereb Blood Flow Metab. 2006 Aug 23

19. Antonetti DA, Barber AJ, Bronson SK, Freeman WM, Gardner TW, Jefferson LS, Kester M,

Kimball SR, Krady JK, LaNoue KF, Norbury CC, Quin PG, Sandirasegarane L, Simpson IA;

JDRF Diabetic Retinopathy Center Group Diabetic Retinopathy: seeing beyond glucose-

induced microvascular disease. Diabetes. 2006 Sep; 55(9):2401-11.

20. Krady JK, Lin HW, Liberto CM, Basu A, Kremlev SG, Levison SW. Ciliary neurotrophic

factor and interleukin-6 differentially activate microglia. J Neurosci Res. 2008 Jan 23;

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