1. Sandra G. Nishikawa
3911 76 Street N.W.
Calgary, AB T3B 2N1
(403) 467-5560 cell
sgnishikawa@gmail.com
snishika@ucalgary.ca
SUMMARY
Techniques routinely used include: isolation of plasmid, phage and genomic DNA from
bacteria, tissue and cell culture sources; plant seeds and leaves; isolation of RNA from tissue
and cell culture sources and subsequent enrichment of m-RNA; synthesis of cDNA; colony,
plaque, Northern and Southern Hybridization; preparation of transformation competent bacterial
cells; transformation of bacterial cells including electroporation and heat shock; isolation of DNA
fragments and cloning of fragments; dideoxy-sequencing of double and single stranded DNA,
thermal cycle sequencing; polymerase chain reaction methodologies including RACE,
INVERSE, RT-PCR, RAPD, nested PCR, qPCR including Taqman and multi-plex qPCR;
restriction analysis of DNA; radio isotopic labelling of DNA and RNA probes, DIG-labelling of
probes; agarose gel electrophoresis ; SDS-PAGE; familiarity of in situ hybridization; chemical
transfection of eukaryotic cells; gene editing using CRISPR-Cas, isolation of PBMC from human
whole blood; maintenance and propagation of a number of cell lines including human, bovine,
murine, mouse embryonic stem cells, human PBMCs and insect; generation of mouse
embryonic fibroblasts; propagation and purification of virus including: Herpesvirus, Reovirus,
Bovine Viral Diarrhea, Hepatitis B and Adenovirus; stimulation and infection experiments with
Hepatitis B and HIV; production of polyclonal and monoclonal antibodies; experience with
number of immunological assays including Western blotting, Immunoprecipitation, RIP, ELIZA,
FITC and FACS; viral neutralization assay, viral plaque titration, radioisotopic labelling of cellular
proteins cloned and expressed in E. coli and Pichia; preparation of bacterial and tissue culture
mediums; animal surgery and routine lab maintenance and management.
PROFESSIONAL EXPERIENCE
May 2014 - Present Research Technician
Dr. Hermann Schaetzl, Universityof Calgary, Dept. of Veterinary Medicine, Calgary
Alberta
Current project involves genome editing using CRSPR-Cas system to knock out the autophagy
gene ATG5 in the neuroblastoma cell line N2a and mouse embryonic fibroblasts (MEF) to
determine its role in Prion clearance and propagation. Knocking out the cellular PRNP gene and
subsequent replacement with CWD PRNP genes.
December 2009 - April 2014 Research Technician

2. Dr. Carla Coffin, Universityof Calgary, Dept. Of Medicine, Immunity, Inflammation and
Infectious Diseases Institute, Calgary, Alberta
Projects include the immunobiology and molecular biology of Hepatitis B virus (HBV)
monoinfection and co-infection with Human Immunodeficiency Virus(HIV) infected patients.
Areas of study are the molecular evolution, host immune responses, and viral reservoirs of HBV
which may lead to a better understanding of the host and virological factors impacting viral
persistence and disease progression. With a better understanding of the underlying
mechanisms of these factors it is hoped to be able to tailor treatment of patients and improve
their clinical outcomes.
November 2002 – December 2009 Research Technician
Dr. Derrick Rancourt, Universityof Calgary, Dept.of Molecular Biology and Biochemistry,
Cancer Research Group, Calgary, Alberta
Projects include the expression of implantation serine proteases in bacteria and yeast.
Implantation serine proteases are thought to be responsible for implantation of embryos in the
uterus of mammals. Was a key member in the development of lambda phage as a vaccine
platform technology. Using this technology it is hoped that vaccines could be made more quickly
and at lower cost. As well, with current threat of bioterrorism it would make a safer vaccine in
that it could not be usurped as a potential weapon. I was also involved in differentiation of
mESC into cartilage and bone as a model for possible use in regenerative medicine.
March 1999 – September 2002 Research Technician
Dr. Patrick Lee, Universityof Calgary, Dept. of Microbiology and Infectious Diseases,
Cancer Research Group, Calgary, Alberta
Projects included the investigation into the use of Reovirus to treat solid tumors. Cell lines were
assessed to determine their susceptibility to infection by Reovirus, as demonstrated through
cytopathic effect, cell labelling western blotting and production of viral progeny. Cell line positive
for susceptibility in vitro were tested in tumor models of nude and SCID mouse. Involved in
determining the role of Reovirus outer capsid protein, sigma 3, in relation to anti-viral cellular
protein PKR. Routine lab maintenance.
December 1998 – March 1999 Research Technician
Dr. Bob Forrester, Agriculture Canada Research Technician, Lethbridge, Alberta
Project involved RAPD (random amplified polymorphic DNA) PCR of DNA isolated from an
anaerobic species of bacteria found in the rumen of cattle.
July 1998 - November 1998 Research Technician
Dr. Sam Lee, Universityof Calgary, Gastrointestinal Research Group, Calgary, Alberta

3. Research in this lab focused on diseases of the liver such a cirrohsis. Rats had their bile ducts
surgically ligated to induce liver disease, urine, blood and tissue samples were biochemically
tested for presence or absence of proteins.
May 1996 – June 1998 Contract Research Associate
Dr. Doug Colwell, Agriculture Canada and Central Biotech, Lethbridge, Alberta
Worked independently on the development of a vaccine against the cattle warble fly,
hypoderma. Cloned three proteins that were isolated from the salivary glands of 1st instar larva.
Cloned proteins were expressed in bacteria and yeast. The proteins were isolated and injected
into cattle as a vaccine. Cattle were then infected with freshly hatched larva. Efficacy was
determined by the number of larva that survived to adulthood as mature flies.
August 1995 - April 1996 Molecular Biology Technician
Dr. Phyllis LaValle, Universityof Calgary, Dept. of Biochemistry, Joint Injury Research
Group, Calgary, Alberta
Work in this lab focus on the differentiation of small chondrocytes to hypertrophic chondrocytes
using molecular and cell biology techniques. The use of differential display was employed to
pick up RNA messages that are common in one but not the other cell type. Messages that were
differentially expressed were chosen as probes in library screening.
November 1994 - July 1995 Molecular Genetics Technician
Dr. Andre LaRoche, Agriculture Canada, Lethbridge, Alberta
Projects involved RAPD (random amplified polymorphic DNA), PCR of single-strand DNA in
wheat species, differential display in canola. Routine lab management.
July 1994 - October 1994 Research Assistant
Dr. Fred Biddle, Universityof Calgary, Dept. of Medical Genetics, Calgary, Alberta
Mapping mouse microsatellites using MapPAIRS (Research Genetics Inc.).
March 1989 - June 1994 Molecular Biology Technician
Dr. Gordon Dixon, Universityof Calgary, Dept. of Medical Biochemistry, Calgary, Alberta
Worked on several projects with limited supervision which allowed me to develop my own
problem solving skills. As a member of the research team I have worked on projects including a
joint venture involving Alta Genetics Inc. and The University of Calgary. This project involved
cloning of male specific sequences from bovine DNA for sex determination of preimplantation
embryos. I was also involved in cloning of H3 histone variants and protamine genes from
salmonid fish.
December 1988 – March 1989 Research Technician
Dr. Saad Masri, Animal Diseases Research Institute, Agriculture Canada, Lethbridge,
Alberta
Experiments involved the cloning of genes from bovine viral diarrhea (BVD) virus.

4. January 1988 – October 1989 Research Technician
Dr. Wally Dixon, Universityof Calgary, Dept. of Oncology, Calgary, Alberta
Involved in the molecular cloning of melanoma associated antigen.
October 1985 – December 1987 Research Technician
Dr. Tony Schryvers, Universityof Calgary, Dept. of Microbiology and Infectious Diseases,
Calgary, Alberta
Worked on the development of monoclonal antibodies against Aspergillus fungus species for
possible diagnostic applications. Also involved the investigation of iron-binding proteins of
Neisseriaceae receptor identification, characterization, cloning of this protein and its regulatory
regions.
January 1985 - August 1985 Research Technician
Dr. Patrick Lee, Universityof Calgary, Dept. of Microbiology and Infectious Diseases,
Calgary, Alberta
This position involved mostly laboratory maintenance.
EDUCATION
B. Sc., 1984 - University of Lethbridge, Lethbridge, Alberta, Canada
PERSONAL PROFILE
I enjoy a variety of activities including motorcycling, camping, home improvement projects and
minor motorcycle repairs. I am a Canadian citizen, single and a very dedicated worker. I have
demonstrated the ability to work without supervision, and I can be an important member of any
team. I am a competent teacher of laboratory methods to trainees. I hope to apply my previous
knowledge while further developing my abilities in my chosen career.
PUBLICATIONS
Lee Zengina, Nishikawa Sandra, Gao Shan Eksteen J. Bertus, Czub Marcus, Gill M. John,
Osiwy Carla, van der Meer Frank, van Marle Guido, Coffin Carla S. Detection of Hepatitis B
Virus (HBV) Genomes and HBV Drug Resistant Variants by Deep Sequencing Analysis of HBV
Genomes in Immune Cell Subsets of HBV mono-infected and/or Human Immunodeficiency
Virus Type-1 and HBV Co-infected Individuals. submitted PLos One
Coffin CS, Osiwy C, Gao S, Nishikawa S, van der Meer F, van Marle Guido. Hepatitis B virus
(HBV) variants fluctuate in paired plasma and peripheral blood mononuclear cells among patient
cohorts during different chronic hepatitis B (CHB) disease phases. J. Viral Hepat. April;
22(4):416-26(2015)

5. Coffin CS, Mulrooney-Cousins PM, Osiowy, van der Meer F, Nishikawa S, Michalak TI, van
Marle G, Gill MJ. Virological characteristics of occult hepatitis B virus in a North American cohort
of Human innumodeficiency virus type 1- positive patients on dual active anti HBV/HIV therapy.
J. Clin. Virology Aug;60(4):347-53
Ding H, Keller KC, Martinez IK, Geransar RM, Zur Nieden KO, Nishikawa SG, Rancourt DE, Zur
Nieden NI. NO/beta-catenin crosstalk modulates primitive streak formation prior to embryonic
stem cell osteogenic differentiation. J. Cell Sci. 2012 Sept. 3
Thomas BS, Nishikawa S, Ito K, Chopra P, Sharma N, Evans DH, Tyrell DH, Bathe OF,
Rancourt DE. Peptide Vaccination is superior to genetic vaccination using a recombineered
bacteria phage subunit vaccine. Vaccine, 30(6):998-1008 (2012)
Sharma N, Kumar R, Renaux B, Saofeddine M, Nishikawa S, Mihara K, Romachandran R,
Hollenberg MD, Rancourt DE. Implantation serine proteinase I exhibits mixed substrate
specificity that silences signaling via proteinase-activated receptors. PLoS One 6(11): e27888
(2011)
Yamashita A, Nishikawa S, Rancourt DE. Identification of five developmental processes during
chondrogenic differentiation of embryonic stem cells. PLoS One, 5(6): e10998 (2010).
Yamashita A, Nishikawa S, Rancourt DE. Microenvironment modulates osteogenic cell lineage
commitment in differentiated embryonic stem cells. PLoS One 5(3): e 9663 (2010)
Thirukkumaran CM, Nodwell MJ, Hirasawa K, Shi ZQ, Diaz R, Luider J, Johnston, RN, Forsyth
PA, Magliocco AM. Lee PWK, Nishikawa S, Donnelly B. Coffey M, Trpkov K, Fonseca K, Spurell
J, Morrison DJ. Oncolytic viral therapy for prostate cancer: efficacy of reovirus as a biological
therapeutic. Cancer Research,70(6) 2435-44 (2010).
Yang WQ, Lun X, Palmer CA, Wilcox ME, Muzik H, Shi ZQ, Dyke R, Coffey M, Thompson B,
Hamilton M, Nishikawa SG, Brasher PM, Fonseca K, George D, Rewcastle NB, Johnston RN,
Stewart D, Lee PW, Senger DL, Forsyth PA. Efficacy and safety evaluation of human reovirus
type 3 in immunocompetent animals: racine and nonhuman primates. Clin. Cancer Research,
10(24) 8561-76: (2004)
Hirasawa K, Nishikawa SG, Norman KL, Coffey MC, Thompson BG, Yoon C-S, Waisman DM,
and Lee PWK. Systemic reovirus therapy of metastatic cancer in immune competent mice.
Cancer Res., 63(2): 348-53(2003)
Alain T, Hirasawa K, Pon KJ, Nishikawa SG, Urbanski SJ, Auer Y, Luider J, Martin A, Johnston
RN, Janowska-Wiezorek A, Lee PWK and Kossakowska A. Reovirus therapy of lymphoid
malignancies. Blood, 100(12):4146-53(2002)
Norman KL, Coffey MC, Hirasawa K, Demetrick DJ, Nishikawa SG, DiFrancesco LM, Strong JE
and Lee PW. Reovirus oncolysis of human breast cancer. Hum. Gene. Ther. 13(5):641-52
(2002).
Hirasawa K, Nishikawa SG, Norman KL, Alain T, Kossakowska A and Lee PW. Oncolytic
reovirus against ovarian and colon cancer. Cancer Res. 62(6):1696-701. (2002).
Wilcox ME, Yang W, Senger D, Rewcastle NB, Morris DG, Brasher PM, Shi ZQ, Johnston RN,
Nishikawa S, Lee PW and Forsyth PA. Reovirus as an oncolytic agent against experimental
human malignant gliomas. J. Natl. Cancer Inst. 93(12):903-12. (2001)

6. Winkfein, R.J., Nishikawa, S., Connor, W. and Dixon, G.H. Characterization of a marsupial
sperm protamine gene and its trasnscripts from the North American opossum (Didelphis
marsupialis). Eur. J. Biochem. 215: 62-72 (1993).
Stros, M., Nishikawa, S. and Dixon, G.H. cDNA sequence and structure of a gene encoding
trout testis High-Mobility-Group-I protein. Eur. J. Biochem. 222: 581-591