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Speaker Proposals
being Accepted!
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Overview
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New to the RNAi Field? Need an Introductory Course?
October 18 2:00-5:30 pm RNAi for Beginners Short Course
This course is designed for bench scientists and managers in both Pharma/Biotech or Academic laboratories who might be considering or have just begun to use RNAi as a
tool for Functional Analysis or Target Validation.
This Course Will Cover:
- An introduction to RNAi, choosing the best reagents to use
- How to set up a high-throughput RNAi library screening laboratory using either chemically
- synthesized or expressed RNAi reagents
- What kind of assays can be run, what automation is required for RNAi library screens
- How to use RNAi effectively in hard to transfect cell lines and in primary cells
- What is the state-of-the-art with regard to the use of RNAi in vivo
- How to make sense of data generated using RNAi reagents (library screens and one-off
experiments)
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Tutors
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Moitreyee Chatterjee-Kishore Ph.D., Principal Scientist/Group Head-Inflammation,
Applied Genomics, Wyeth Research |
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Vivek Mittal, Ph.D., Assistant Professor, Cancer Genome Research Center,
Cold Spring Harbor Laboratory |
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Thijn Brummelkamp, Ph.D., Fellow,
Whitehead Institute
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Picture not available |
Craig S. Mickanin, Ph.D., Senior Scientist II,
Novartis Institutes for BioMedical Research
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Wednesday, October 19
7:00-8:30 Registration
| 7:30-8:15 Breakfast Workshop
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Transitioning RNAi from a Research Reagent to a Validated Tool
Mr. Tobias Bergauer, Cell Biology, QIAGEN GmbH
The application of RNAi to target validation has the potential to revolutionize gene fictionalization. Increasingly, researchers are interested in developing the ability to perform large, genome-wide screens using
RNAi. The flexibility, reliability, and performance of QIAGEN-designed siRNA make it the reagent of choice for high-throughput screening approaches for target validation. An integrated system for gene knockdown and gene expression analysis has been developed by
QIAGEN, utilizing genome-wide siRNA and gene expression assays to transform RNAi research reagents into standardized scientific tools. |
Sponsored By
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In vivo RNAi for Functional Analysis and Target Validation
8:30-8:40 Chairpersons's Remarks
John F. Cryan, Ph.D., Laboratory Head, Neuroscience Research, Novartis Institutes for BioMedical Research, Novartis Pharma AG
Oncology Case Study
8:40-9:10 RNAi for Pharmaceutical Target Validation in Animal Models
Hans Winkler, Ph.D., Director, Functional Genomics, Johnson & Johnson Pharmaceutical R&D
RNAi is a natural mechanism that has been exploited to repress gene expression in many biological systems. Naturally, the ability to target specifically the inhibition of gene expression in animals is of great value for target validation in the pharmaceutical drug discovery process. We have used a lacZ expressing mouse strain in conjunction with
lacZ-specific siRNA to assess various delivery systems and application routes for siRNA application
in vivo.
Oncology Case Study
9:10-9:40 Identification and Validation of Novel Drug Targets Using RNAi
Dr. Lata Jayaraman, Research Investigator, Oncology Drug Discovery, Bristol Myers Squibb Co
The biopharmaceutical industry today faces a major challenge in how best to identify and select the next generation of molecular targets for oncology. An impressive array of potential new cellular targets, suitable for therapeutic intervention, has been revealed by the recent completion of the human genome sequencing project. Approaches as varied as transcription profiling, proteomics and the use of small interfering RNAs are all being exploited in the race to select the most promising candidate drug targets. We have used genome-wide functional RNAi screens in model organisms to mine key oncogenic or tumor suppressor pathways as a viable approach to identifying novel targets for oncology drug discovery. This presentation will highlight the advantages and challenges associated with such an approach.
Oncology Case Study
9:40-10:10 Evaluating HIF-1a as a Cancer Therapeutic Target via Inducible RNAi in vivo
Yu Shen, Ph.D., Associate Research Investigator, Cancer Research, Abbott Laboratories
Validating potential targets is one of the most critical steps in drug discovery. In this study, we tested the feasibility of using inducible RNAi
in vivo to obtain an unbiased evaluation on the efficacy of inhibiting HIF-1a in established tumors. We demonstrated that HIF-1a inhibition resulted in transient tumor regression or stasis, and inhibiting HIF-1a in early stage tumors was found to be more efficacious than inhibiting HIF-1a in more established tumors. A differential requirement of HIF-1a for tumor growth was also observed among different tumor types. Examination of tumors resistant to HIF-1a inhibition suggested that the resistance might result from a less hypoxic tumor environment and the level of HIF-1a expression in tumors may be a useful marker for predicting tumor response to HIF-1 inhibition. This study demonstrated that inducible RNAi is a versatile tool for evaluating cancer targets
in vivo. In addition to broad implications on in vivo validation of cancer targets, results from this study will also be instructive for practical applications of HIF-1-based cancer therapeutics. In addition to the inducible xenograft tumor model, the creation of
RNAi-based knockdown animals and the direct use of chemically synthesized siRNA for
in vivo applications will also be discussed.
| 10:10-11:00 Coffee Break, Poster and Exhibit Viewing |
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CNS Case Study
11:00-11:30 Use of RNAi in Target Validation for Neuropsychiatric Disorders
John F. Cryan, Ph.D.
The burgeoning use of gene expression analysis to detect potential targets relevant to psychiatric and
neurological disorders necessitates the validation of such targets in vivo. By virtue of its ability to rapidly
silence genes even in adult animals, RNAi has recently emerged as a superior alternative to the
traditional gene-silencing approach for target validation. Efforts to achieve widespread gene knockdown
for target validation in the adult brain will be discussed.
CNS Case Study
11:30-11:45 Silencing Mutant SOD1 using RNAi Protects against Neurodegeneration and Extends Survival in an ALS Model
Scott Ralph, Ph.D., Senior Scientist, Neurobiology, Oxford Biomedica Ltd.
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Amyotrophic Lateral Schlerosis
(ALS) is a fatal neurodegenerative disease causing the selective death of motor neurons of the brain stem and spinal cord. Some familial forms of ALS are caused by dominantly inherited mutations in the gene encoding superoxide dismutase (SOD1). We have designed lentiviral vectors to express functional RNAi molecules targeted against human SOD1 and demonstrated that delivery of this vector to vulnerable motor neuron populations of transgenic mice carrying a copy of the mutant human SOD1 gene, prevents
neurodegeneration, delays the onset of ALS symptoms and significantly increases the lifespan of these animals. |
11:45-12:15 Target Discovery through Whole Genome RNAi-Based Screens in Cells
Hao Li, Ph.D., Laboratory Head, Department of Functional Genomics, Novartis Institutes for Biomedical Research
RNAi technology has provided a powerful reverse genetic approach for systematic analysis of gene function and rapid identification of genes involved in specific cellular processes and pathways. We are currently carrying out whole genome dsRNAi screens using the fruitfly Drosophila melanogaster as a model system. Long RNAi molecules (~500bp) are efficiently taken up by a variety of Drosophila cell types even in the absence of transfection reagents, and the resulting intracellular production of small RNAs leads to highly efficient and gene-specific reductions in gene function. Given the relatively low genetic redundancy in
Drosophila, the established functional conservation between human and Drosophila for many disease-relevant pathways and processes, we expect that these efforts will complement and add value to similar approaches being taken in mammalian cells. We will report results from our genome-wide screens carried out to identify genes modulating several signaling pathways and cellar processes.
12:15-12:35 In vivo
RNAi: Where Are We Going?
Panelists: All above speakers
| 12:35-1:45 Lunch and Learn Workshop
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Sponsored by
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Increased Potency of 27mer siRNA Duplexes
Mark A. Behlke, Ph.D., Vice President of Molecular Genetics & Biophysics, Molecular Genetics, Integrated DNA Technologies
In attempts to identify RNAi triggers that effectively function at lower concentrations, we found that synthetic RNA duplexes of ~27 nucleotides in length can be up to 100 fold more potent than corresponding 21mer
siRNAs. The enhanced potency of the longer duplexes is attributed to the fact that they are Dicer substrates, directly linking the production of siRNAs to incorporation into RISC. Not all 27mers show this increased potency, mostly due to variation in dicing patterns. We have defined methods to direct dicing to produce specific, desired products and thereby make design of high-potency 27mer siRNAs more predictable. Unexpectedly, we observe that dicing introduces functional asymmetry into the potency of an RNA duplex independent of sequence. This effect is directional and influences strand loading into RISC. |
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Genome-Wide RNAi Screens
1:45-1:50 Chairperson’s Remarks
Dr. Kader Thiam, Director of Transgenic Technologies, genOway |
Sponsored by |
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1:50-2:20 RNAi: A Key Tool to Investigate Novel Genes from Genetics/Genomics Screens
DR.Cristina Rondinone, Director, Research Metabolic Diseases, F. Hoffmann-La Roche
Inc.
Interrogation of cellular process using RNAi holds extraordinary promise for the identification of novel mediators of signal transduction pathways and potential drug targets. RNAi can be a useful tool to increase the biological understanding and infer the function of novel protein targets that can emerge from genetic linkage studies and genomics. This talk will discuss how we are using RNAi to test this hypothesis, and validate, select and prioritize the best targets for metabolic diseases.
2:20-2:50 High-Throughput Screening Using Genome-Wide siRNA Libraries
Zicai Liang, Ph.D., Associate Professor, Center for Genomics and Bioinformatics, Karolinska Institute, Stockholm
A random siRNA library and a library of pre-designed siRNA covering the whole human genome have been created using an opposing-dual promoter system. The libraries have been used to carry out phenotype-driven screening of novel functional genes with promising results. This talk will cover the creation of the vector system, the libraries, the screening process and results as well as off-target considerations.
2:50-3:20 RNAi Library Screens
Moitreyee Chatterjee-Kishore, Ph.D., Principal Scientist / Group Head-Inflammation, Applied Genomics, Biological Technologies
Abstract Unavailable at Time of Printing
3:20-4:00 Refreshment Break, Poster and Exhibit Viewing
Predicting and Avoiding Off-Target Effects
4:00-4:30 Designer siRNAs and Pitfalls in a Stolen Natural Process
Sumedha Jayasena, Ph.D., Senior Principal Scientist, Oncology Research, Amgen Inc.
Since their inception, siRNA molecules have been used to silence a vast number of genes in tissue culture cell lines, albeit with limited
in vivo applications. At least in vitro, siRNA-mediated gene silencing has already become a mainstay in target discovery, pathway analysis and therapeutic development. However, with more and more applications, a number of issues has been discovered to challenge the specificity of this approach. This presentation highlights some approaches to overcome such challenges.
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4:30-5:00 siRNA Off Target Effects: Proposed Mechanism and Strategies to Minimize the Phenotypic Impact
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Sponsored by |
Anastasia Khvorova, Ph.D., Director of Biology, Biology R&D, Dharmacon, Inc.
RNAi is a remarkable tool for gene function analysis, pathway elucidation, drug target validation, and gene mapping. While RNAi is typically associated with unparalleled specificity in gene silencing studies, two major issues include (1) understanding and predicting the biological impact of siRNA induced off-target effects and (2) strategies that help minimize off-target-induced phenotypes. Reducing off-target events is paramount for
RNAi-based large-scale screens to reduce the rate of false-positives and unnecessary follow-up confirmatory experiments. As partial sequence identity between a given siRNA and corresponding target site is sufficient to induce cleavage, siRNA specificity is currently viewed as a major technological challenge. This talk will discuss:
• Proposed mechanisms underlying off-target effects,
• Rules that aid in recognizing potential off-target events,
• Examples of the functional and phenotypic consequences of off-target events in high throughput screens, and
• Methods to mitigate off-target events and enhance specificity including pooling functional siRNAs into single potent reagents and the application of novel chemical modification patterns. |
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5:00-5:15 Advances in Stealth™ RNAi and miRNA Design for Highly Effective Gene Silencing
Michaeline Bunting, Ph.D., Senior Scientist, RNAi Research Area Manager, RNAi Research, Invitrogen Corporation
Recent advances in the design of Stealth™ RNAi and miRNA expression vectors for highly effective and specific gene silencing will be presented. Stealth™
RNAi, a chemically modified synthetic dsRNA, was developed to achieve high specificity and greater stability compared to unmodified siRNA molecules. Using computational analysis of large sets of functionally active Stealth™ RNAi molecules, we have developed and validated a highly predictive algorithm to identify potent Stealth™ RNAi target sequences. We will also present the construction of Pol II-driven
miRNA-based RNAi vectors to achieve tissue specific inhibition of multiple targets from a single vector. These vectors express artificial miRNAs which are designed to perfectly complement, and therefore cleave, endogenous target transcripts with a high rate of success. Use of this system to target endogenous genes, including through lentiviral delivery, will be
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Sponsored by: |
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5:15-5:45 Panel
Design is Critical to a Successful RNAi Experiment – From Off-Target Effects and Beyond
Panelists:
Cristina M. Rondinone, F. Hoffman-La Roche Ltd.
Michaeline Bunting, Invitrogen Corporation
Yu Shen, Abbott Laboratories
Lata Jayaraman, Bristol-Myers Squibb Co.
Hao Li, Ph.D., Laboratory Head, Department of Functional Genomics, Novartis
Institutes for Biomedical Research
Discussion Topics:
• Figuring out which "hits" are actually real following siRNA screening
• Redundancy of family members - a problem to be tackled
• Importance of secondary assays
• Differentiation of stress/tox versus “real” hits
• Handling the data flood generated by high-content screening
Thursday, October 20
8:00-8:30 Registration and Morning Coffee
High-Content RNAi Screening: A Powerful Tool for Target Discovery and Validation
8:30-8:40 Chairperson's Remarks
Dr. Edward Weinstein, Principal R&D Scientist,
Functional Genomics, Sigma-Aldrich Corporation
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Sponsored by |
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8:40-9:10 Anticancer Drug Development using High-Content Screening and RNAi
Steven Haney, Ph.D., Senior Scientist, Genomics, Wyeth Research
The current focus of target selection is to provide greater success in drug discovery through the identification of better targets. RNAi and High-Content Screening are two technologies that have high potentials to improve target selection, and can be integrated into a common platform that allows target ID, validation and assay development to be closely linked processes. These technologies are quite flexible, allowing for a wide range of cell models to be used for drug discovery, as well as to allow for insights from current clinical experience to be incorporated into the early part of the drug discovery pipeline.
9:10-9:40 RNAi-Based High Content Screen for Oncology Target Discovery and Validation
Judith Wardwell-Swanson, Ph.D., Senior Research Investigator II, Applied
Genomics, Bristol-Myers Squibb Co.
A medium-throughput, ultra-high content approach to the discovery of new drug targets for Oncology will be discussed. Potential drug target candidates first identified from model organism genetic screens were functionally validated in human cell models using
RNAi-based high content assays. The resulting rich datasets were subjected to rigorous data analysis and ultimately used to generate functional response profiles for each of the candidate genes. Using this approach novel drug targets can be rapidly identified, functionally validated and classified within the context of relevant disease models.
9:40-10:10
High-Content RNAi Screening for
Genome-Wide Functional Analysis of Human Kinases in Endocytosis.
Dr. Eberhard Krausz, HT-Technology Development
Studio (TDS), Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)
Although kinases are prominent drug targets, only a
small number has been associated with particular diseases and therefore been
addressed by drug development. However, the availability of RNA interference now
provides the opportunity to gain functional information on each individual
target. We applied a kinase siRNA library to multi-parametric high-content
assays using automated high-throughput microscopy. First, we characterized the
library in a time-course for the impact of target mRNA down-regulation on cell
viability and apoptosis to define a screening window. Subsequently, the library
was applied to primary screens for viral infection; in secondary assays the
pathways were dissected to characterize novel targets in regulating endocytosis.
| 10:10-11:10 Coffee Break, Poster and Exhibit Viewing
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Sponsored by |
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RNAi Delivery: Primary Cells, Difficult to Transfect Cell Lines and More
Chairperson’s Remarks
Christophe Echeverri, Ph.D., CEO/CSO, Cenix BioScience GmbH
11:10-11:40 Lessons Learned from Major Disease-Focused Target Discovery Screens using HT-RNAi and High-Content Microscopy Assays in Human Cells
Christophe Echeverri, Ph.D.
We have carried out HT-RNAi screens using multiplexed, high content microscopy assays in primary and transformed human cells to identify genes involved in several key disease-relevant processes. The screens applied 3 siRNA molecules used individually to target each of ~5,200 genes chosen for their high therapeutic potential. The multiplexing of microscopy readouts, combined with advanced automated image analysis, enabled the generation of rich phenotypic profiles to classify genes. Results and lessons learned will be discussed at the meeting.
11:40-12:10 The Next Frontier in RNAi
Technology: In vivo Relevance
Norbert Perrimon, Ph.D., Department of Genetics, Harvard Medical School,
Howard Hughes Medical Institute
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RNAi screens in tissue culture are attractive because they can be conducted relatively fast and
also have technical features (i..e., multiplexing, subcellular resolution) that are not easily
achievable in vivo. However, we have to be cautious that the findings made in culture are meaningful
in vivo. I will discuss RNAi screens in primary cells, whose differentiation programs follow
closely their in vivo differentiation, as well as our effort at developing genome-wide transgenic
RNAi methods.
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12:10-12:25 Collaborative Approach to Using siRNA in the Discovery of Disease-Modifying Drug Targets
Dr Richard Janssen, Senior Scientist, Galapagos NV
Our integrated approach to target discovery starts with genome-wide RNAi screens in disease relevant human primary cells, followed by more complex
in vitro and ex vivo assays - all of which functionally confirm a target's role in the disease pathology. In collaboration with major pharmaceutical companies, we have implemented this technology platform across a wide range of disease areas to deliver targets into compound screening. Such a technique provides a practical framework for using siRNA to generate novel targets - potentially leading to the discovery of new therapeutics.
12:25-12:30 Chairperson's Wrap-Up
12:30-2:00 Luncheon in the Exhibit Hall
(Last Chance for Poster and Exhibit Viewing)
2:00-3:15 Buzz Sessions - Roundtable Breakout Discussion
Ice Cream Served
Table Topics:
• In vivo RNAi Delivery
• Validation of RNAi knockdown
• Deciphering the results of genome-wide RNAi screens
• Primary Cell Transfection-Hosted by Dr. Titus Kretzschmar, Vice President R&D,
amaxa GMBH
• RNAi and High-Content Imaging
• Cancer Targets Validation
• Valid Disease Models
• Cell-Based Chemical Biology
• Positioning the Value of Enabling Technology
• The Importance of Moving Downstream with Enabling Technology
• Measuring the Financial Impact of Technology |
3:15-3:20 Chairperson's Remarks
Christophe Echeverri, Ph.D., CEO/CSO, Cenix BioScience GmbH
3:20-3:50 Laser-Mediated Delivery of siRNA Into Difficult Cell Types with High-Efficiency and Low Toxicity
Fred Koller, Ph.D., President and Chief Technology Officer, Cyntellect
Efficient delivery of siRNA into a variety of cell types is essential in many fields including basic research, applied drug discovery, and clinical applications. Unfortunately, current methods are limited by the types of cells that can be transfected, often due to poor efficiency, cell toxicity, and/or physiological changes that are induced by the transfection method. To address these issues, a novel laser-based cell transfection approach was developed. High-efficiency, laser-mediated delivery of siRNA was demonstrated in a number of recalcitrant cell types (e.g., human B- and T-cells, neurons), in each case with less cell toxicity than with standard methods. Laser-mediated transfection of siRNA was shown to be consistent across many cell types, enabling RNAi studies to be conducted in
difficult-to-transfect cells.
3:50-4:20 Hydrodynamic Delivery of siRNA to Regulate Exogenous and Endogenous Genes Expressed by Hepatocytes
Qiuming Chu, Ph.D., Staff Scientist, Applied Discovery Research, Genzyme
Delivery of small interfering RNAs (siRNAs) to target cells in vivo is a key obstacle to studying their function in rodents. Hydrodynamic delivery has been shown to be a very powerful method to deliver nucleic acids to rodent liver, and in particular, hepatocytes, which represent an attractive target for studying siRNA function
in vivo. Here, the function of four siRNAs will be described following hydrodynamic delivery to the mouse liver. Two siRNAs are against exogenous transgenes transferred into hepatocytes, namely, chloramphenicol acetyl transferase and human a-galactosidase A. Two additional siRNAs are against endogenous genes normally expressed by hepatocytes, namely, acetyl-CoA carboxylase 2 and acid sphingomyelinase. In addition, possible methods or conditions for hydrodynamic delivery of siRNAs to the rat liver and for hydrodynamic delivery of nucleic acids in rabbits with balloon catheters will be discussed.
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4:20-5:30 Novel Technology Approaches to Delivery
This session will feature a series of four short commentaries highlighting emerging delivery techniques and methods.
RNAi Screening in vitro: High-throughput Delivery Methods and siRNA Library Applications
Dr. David Dorris, Dir RNAI Technologies, RNAI Technologies, Ambion Inc.
RNAi has been rapidly adopted for functional genomics, pathway analysis, and drug target validation, and is now being used in genome scale RNAi screens to discover and characterize gene functions directly in human cells. I will discuss high throughput siRNA delivery methods for both immortalized cell lines and primary cell types, as well as successful applications of siRNA libraries.
Using RNAi to Study Gene Function in Liver
David L. Lewis, Ph.D., Program Director - RNA Interference, Mirus Bio Corporation
The ability to use RNAi in mammalian models would greatly aid in identifying novel drug targets and eliminate those targets that are not disease-relevant. However, delivery of siRNAs or shRNA expression vectors in vivo can be technically challenging. We have found that hydrodynamic intravascular injection allows for simple and efficient delivery of naked siRNA in a variety of mammalian species. Using this method, we examined the effects of knocking down a hepatocyte-specific target gene in mouse liver. Transcriptome profiling of the siRNA treated mice reveals a phenotype related to that of knockout or agonist treated animals. Delivery of siRNA and shRNA expression vectors to liver using systemically administered nanoparticles will also be discussed.
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RNAi SmartReport Includes:
• 84 companies
• 146 press releases issued during the last twelve months.
Special offer for attendees!
Report Price Only $325
For
Sponsorship and Exhibit Opportunities
John Yurewicz, Manager of Business Development
Phone: 617.630.1383, Cell: 857-636-8188 • Email: jyurewicz@healthtech.com
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