Work in Progress
My current research program is focused on:
- Monitoring and detection of biological threats
- Diagnosis of infectious diseases and cancer
- Targeting of tumor cells
To attack these problems, I employ methods of Directed Molecular Evolution, Multivalent Phage Display
and Phage Nanobiotechnology that have been developed by my group. This program is a continuation of
my previous study of the Combinatorial Biochemistry and Landscape phage Libraries.
I began my research activity in 1969 as a student at the
Moscow State University in the
Laboratory of Phosphor-organic Chemistry headed by
Prof. E.E. Nifant'ev.
My goal was to synthesize molecules that could normally operate in living cells. In my master diploma project,
I developed new phosphorylation agents for synthesis of DNA, glycosyl-phosphates and other important natural
compounds. After graduating from the University in 1972, I continued my research as a Ph.D. student in
the Institute of Organic Chemistry, USSR Academy of Sciences,
Moscow. My supervisors were
Academician N.K. Kochetkov and
Prof. V.N. Shibaev — internationally recognized leaders in DNA and carbohydrate chemistry. In
my Ph.D. dissertation project, I studied biosynthesis of Salmonella antigens using synthetic analogs of
thymidine-diphosphate-rhamnose. Results of this study were presented in 7 papers.
Since 1977, I focused my scientific interests on directed molecular evolution, synthesis and designed
reconstruction of DNA, phage display, protein drug design, and diagnostics.
The 70th revolutionized biotechnology by inventing recombinant DNA.
Thus, since 1977, after establishing my laboratory in Scientific Association “Vector”, Novosibirsk
(USSR), I focused my scientific interests on directed molecular evolution,
designed reconstruction of DNA, phage display, protein drug design, and
diagnostics. As result, my group developed first analogues of oligonucleotides
inducing site-directed mutations in vitro and in vivo
- Petrenko V.A., P.I. Pozdnyakov, S.M. Kipriyanov, A.N. Boldyrev, L.N. Semenova,
G.F. Sivolobova (1986) Site-localized mutagenesis directed by phosphotriester
analogues of oligonucleotides. Bioorganicheskaya khimiya., 12, 289-292. Soviet
Journal of Bioorganic Chemistry Engl. Tr., 1986, 12, 586-597.
- Petrenko VA, Kuzmicheva GA, Tatkov SI, Krasnoborov II, Ilichev AA, Drutsa VL,
Purmal AA, Shabarova ZA. Study of the mutagenic properties
of oligonucleotides containing and internucleotide pyrophosphate bond using a
mutation system based on phage M13. Mol Biol (Mosk). 1991 Nov-Dec; 25(6):1539-45
- Petrenko V.A., S.M. Kipriyanov, A.N. Boldyrev, P.I. Pozdnyakov (1989).
Mutagenesis directed by phosphotriester analogs of
oligonucleotides: a way to site-specific mutagenesis in vivo. Mol Gen Mikrobiol Virusol. 1989 Mar;(3):35-9.
- Petrenko V.A., L.I. Karpenko, S.I. Tatkov, O.Yu. Smirnova, G.A. Kuzmicheva,
A.A. Ilyichev (1991) Mutagenesis induced by oligonucleotide phosphotriester
analogs and directed at double-stranded DNA breaks. Molekulyarnaya Genetika
Mikrobiologia Virusologia., 10, 19-20.
We inventored a method allowing to introduce mutations localized in the
operator-promotor regions of genome:
- Gusev V.A., V.A. Karginov, V.V. Kravchenko, V.A. Petrenko, N.A. Petrov, L.N.
Semenova (1981) Modification of the operator-promoter site of lambda phage DNA
in the specific complex with E.coli RNA polimerase. Doklady Akademii nauk SSSR.,
Minenkova O.O., A.A. Ilyichev, G.P. Kishchenko and V.A. Petrenko (1993) Design
of specific immunogen using filamentous phages as a carrier. Gene, 128, 85-88.
Multivalent Phage Display
In 1985, George Smith published in
Science his revolutionary paper describing cloning of foreign gene as a fusion to
the Minor coat protein pIII of the filamentous phage fd (Times Cited: 1,971). Later
this technique was named “Phage Display”:
- Smith G.P. and Petrenko V.A. (1997) Phage Display. Chemical Reviews, V.97, N2,
After Smith’s publication, I was fascinated to learn if filamentous phage can tolerate multiple mutations and
insertions of foreign peptides into the Major coat protein pVIII, 2,700 copies of which cover the
viral DNA to form viral capsid. It was commonly believed that pVIII is extremely conservative and cannot
be modified. Using methods of site-directed mutagenesis, we constructed vectors
that allowed us to check this hypothesis.Our efforts resulted in construction of the first recombinant phages
with the pVIII protein fused to foreign peptides:
- Ilyichev A.A., O.O. Minenkova, S.I. Tatkov, N.N. Karpyshev, A.M. Eroshkin, V.A.
Petrenko, L.S. Sandakhchiev (1989) Production of a viable variant of phage M13
with incorporated foreign peptide in the major coat protein. Doklady Akademii
nauk SSSR., 307, 481-483. Doklady Biochemistry (Proc. Acad. Sci. USSR)-Engl.Tr.,
1989, 307, 196-198.
- Kishchenko G.P., O.O. Minenkova, A.A. Ilyichev, A.D. Gruzdev, V.A. Petrenko
(1991) Study of the structure of phage-M13 virions containing chimeric B-protein
molecules. Molekulyarnaya biologiya., 25, 1497-1503. Molecular Biology, 1991,
Inventors of pVIII phage display. Novosibirsk, Russia
(left to right): Gregory Kishchenko, Sergey Tatkov,
Alexander Il’ichev, Galina Kuzmicheva, Valery Petrenko, Olga Minenkova
These preliminary results inspired me to study filamentous phage with pVIII fusions (later named
“landscape phage”) as novel engineered nanomaterials for design of synthetic
vaccines, diagnostics, detectors and drug/gene delivery vectors:
Petrenko V.A., Smith G.P., Gong X. and Quinn T. (1996) A library of organic
landscapes on filamentous phage. Protein Engineering, V.9, N9, p. 797-801
and Smith G.P. (2000) Phage from landscape libraries as substitute antibodies.
Protein Engineering, V.13, N8, p.101-104.
V.A., Smith G.P., Mazooji M.M. and Quinn T. (2002) Alfa-helically constrained
phage display library. Protein Engineering, V.15, No.11, pp.943-950.
and V.J. Vodyanoy (2003) Phage display for detection of biological threat
agents. The Journal of Microbiological Methods, 53/2 pp. 243-252.
D.D. Williams, I.B. Sorokulova, V. Nanduri, I-H. Chen, C.L. Turnbough, Jr. and
V.A. Petrenko (2004).
probes for Bacillus anthracis spores selected from a landscape phage library.
Clinical Chemistry V.50, No.11, p.1899-1906.
biosensor designers (20030 left to right: Jennifer Brigati, Iryna
Sorokulova, I-Hsuan Chen, Eric Olsen, Jane Mount, Valery Petrenko, Vishu Nanduri,
(2008). Landscape phage as a molecular recognition interface for detection
devices. Review. Microelectronics Journal. Vol 39/2 pp 202-207.
Kuzmicheva, P.K. Jayanna, I.B. Sorokulova and V.A. Petrenko (2009) Diversity and
censoring of landscape phage libraries. Protein Engineering, Design, and
Selection. 2009 Jan;22(1):9-18.
Nanobiotechnology. (Valery A. Petrenko and George P. Smith Eds.) RSCPublishing,
My current interest lies in the area of cancer-targeted drug development.
We first demonstrated that cancer-targeted major coat proteins isolated from preselected landscape phages
can be complexed with liposomes, micelles, nanotubes and siRNA, increasing their therapeutic effect:
Jayanna P.K., Torchilin V.P. and Petrenko V.A. (2009) Liposomes Targeted
by Phage Fusion Proteins.
Nanomedicine: Nanotechnology, Biology and Medicine Volume 5, Issue 1, March
2009, Pages 83-89. Epub 2008 Oct 1.
Yang S, Petrenko VA, Torchilin VP. Cytoplasmic Delivery of Liposomes into MCF-7
Breast Cancer Cells Mediated by Cell-Specific Phage Fusion Coat Protein. Mol
Pharm. 2010 Aug 2;7(4):1149-58.
Petrenko VA, Torchilin VP. Paclitaxel-Loaded Polymeric Micelles Modified with
MCF-7 Cell-Specific Phage Protein: Enhanced Binding to Target Cancer Cells and
Increased Cytotoxicity. Mol Pharm. 2010 Aug 2;7(4):1007-14.
PK, Bedi D, Gillespie JW, Deinnocentes P, Wang T, Torchilin VP, Bird RC,
Petrenko VA. Landscape phage fusion protein-mediated targeting of nanomedicines
enhances their prostate tumor cell association and cytotoxic efficiency.
Nanomedicine: Nanotechnology, Biology, and Medicine. 2010
Tiziana Musacchio, Olusegun A. Fagbohun, Patricia
Deinnocentesa, R. Curtis Bird,
Vladimir P. Torchilin, and Valery A. Petrenko. Delivery of
siRNA into breast cancer cells via phage fusion protein-targeted liposomes.
Nanomedicine: Nanotechnology, Biology, and Medicine, 7 (3): 315-323 JUN 2011.
A. Fagbohun, Deepa Bedi, Natalia I. Grabchenko, Patricia A. Deinnocentes,
Richard C. Bird, and Valery A. Petrenko. Landscape phages and their fusion
proteins targeted to breast cancer cells. Protein Engineering, Design &
Selection, 25(6), pp. 271-83, 2012.
Gillespie JW, Petrenko VA Jr, Ebner A, Leitner M, Hinterdorfer P, Petrenko VA.
Targeted Delivery of siRNA into Breast Cancer Cells via Phage Fusion Proteins.
Mol Pharm. 2013 Feb 4;10(2):551-9.
Epub 2013 Jan 8.
Cancer fighters (2011) left
to right: James Gillespie, Valery Petrenko, Tiffani O'Dell, Vasily Petrenko,
Deepa Bedi, Amanda Gross, Lixia Wei.
Cancer Fighters (2013) left to right:
Amanda Gross, James Diskin, Tiffani O'Dell, Clifford Deerman, Logan Stallings,
James Gillespie,Deepa Bedi, Valery Petrenko.
Petrenko VA, Jayanna PK. Phage protein-targeted cancer nanomedicines. FEBS Lett.
2014 Jan 21;588(2):341-9.
Bedi D, Gillespie JW, Petrenko VA. Selection of pancreatic cancer cell-binding
landscape phages and their use in development of anticancer nanomedicines.
Protein Eng Des Sel. 2014 Jul;27(7):235-43.
U.S. Patent 8,137,693 B2. Drug Delivery Nanocarriers Targeted by Landscape
Phage. Valery A. Petrenko; Date of Patent: Mar.20, 2012. Application No.
11/536,844; Filed Sep.29, 2006; Provisional appln. No 60/722,320, filed on
fighters (2015) left to right: Amanda Gross, Logan Stallings, Christopher
Ramhold, Valery Petrenko, Anatoliy Puzyrev, James Gillespie.
As a faculty member in the Department of Pathobiology (since October 2000),
I established the extramurally funded Program in the areas of Directed Molecular Evolution and Targeting. The research program includes:
Construction of new phage vectors and diverse phage display libraries.
Selection of tumor-targeting phage probes and their use for diagnosis and
targeted gene and drug delivery and treatment of cancer diseases.
Selection and evolutionary improvement of phage-derived probes against Salmonella typhimurium,
Bacillus anthracis and other food and environmental threat agents.
Genetically engineered self-assembling of bio-selective layers in biosensors for detection of microbial and toxic threat agents.
Optimization of monitoring and detection of Salmonella typhimurium,
Bacillus anthracis and other threat agents by phage-derived probes
immobilized on the surface of biosensors.