Analysis of the Threat of 

Genetically Modified Organisms 

for 

Biological Warfare 

Jerry Warner, James Ramsbotham, Ewelina Tunia and James J. Valdes 

Center for Technology and National Security Policy 

National Defense University 

May 2011 
2

The views expressed in this article are those of the authors and do not reflect the official policy 

or position of the National Defense University, the Department of Defense or the U.S. 

Government. All information and sources for this paper were drawn from unclassified materials. 

COL (Ret) Jerry Warner is a 1976 graduate of the United State Military Academy at West 

Point and holds a Master of Science degree in “Operations Research and Systems Analysis” from 

the Naval Postgraduate School, and Master of Science degrees in “National Security Strategy” 

and “Information Strategies Concentration Program” from the National War College. He served 

as an Army combat officer in many theatres of operation and his last military assignment was at 

the Office of the Secretary of Defense, Net Assessment. He is currently Managing Director of 

Defense Life Sciences, LLC. 

Alan J. Ramsbotham, Jr., has specialized in advanced technology assessment and national 

security analysis for the past thirty years. Prior to that, as a Navy civil service employee, he held 

a series of responsible engineering research and acquisition management positions. Mr. 

Ramsbotham holds a Masters degree in electronic and electrical engineering from the University 

of Maryland. In his capacity as President of Orion Enterprises, Inc., he has been the principal 

investigator and author of over a hundred assessments or technical papers, including technology 

security assessments in the specific areas of battlefield biotechnology and genomics done for the 

Army Materiel Command. 

Ewelina Tunia is a Research Assistant at the National Defense University Center for Technology 

and National Security Policy. She earned her Masters degree in Security Studies from Georgetown 

University’s Edmund A. Walsh School of Foreign Service and Bachelors degree in Political Science 

from Hunter College, City University of New York. Previously, she held internships at Amnesty 

International and the United Nations and was an intelligence analyst for the U.S. Army National 

Guard. 

James J. Valdes is a Senior Research Fellow at the National Defense University’s Center for 

Technology and National Security Policy and the Army’s Scientific Advisor for Biotechnology. 

Dr. Valdes received a PhD in neuroscience from Texas Christian University and was a 

postdoctoral fellow at the Johns Hopkins Medical Institutes. He has published more than 120 

papers in scientific journals and was a 2003 Presidential Rank Award winner. 
3

EXECUTIVE SUMMARY

Evaluating the potential threats posed by advances in biotechnology, especially genetically 

modified organisms (GMOs) and synthetic biology remains a contentious issue. Some believe 

that, inevitably, these advances will lead to a catastrophic biological attack. Others believe that, 

despite these advances, the scientific and technical requirements, as well as the fundamental laws 

of natural selection will prevent such an attack. 

To better understand this issue, this study narrowed the scope of consideration in several 

dimensions. First, our analysis primarily focused on what we defined as a “catastrophic 

biological attack”, with a required level of damage more associated with biological warfare than 

bioterrorism.1 This damage would need to be direct in nature where the effect is more physical 

than psychological. Second, this biological attack would be restricted to the United States, not 

another nation or entity. In this sense, U.S. geography, climatology, infrastructure and medical 

systems play to counterbalance any potential biological attack. Even within a more narrow 

scope, there remains inherent complexity and uncertainty which, combined with the considerable 

rate of change for biotechnology, defies a simple, straightforward answer. 

We approached the issue by establishing an “Analytical Framework”—a baseline of the technical 

requirements to “play” in the field of GMOs at the scale of biological warfare. The primary focus 

of the framework are those aspects of the technology directly affecting humans by inducing 

virulent infectious disease, or through expression of toxins or suppression of the immune 

response of target subjects. Parallel threats exist for animals and plants in the food chain and, 

secondarily, in the ecosphere. Although not specifically included in this analysis, those threats 

can also be evaluated within the analytical framework. To establish our analytical framework, we 

focused on the engineering of novel single-cell microorganisms previously unknown in nature as 

described by four conditions: 

1. Modification of known pathogen microorganisms to new functionalities 

 2. Modification of nonpathogens to become pathogenic 

 3. Synthesizing pathogenic microorganisms de novo

 4. Synthesizing completely artificial or “abiotic” pathogenic “cells” or biomolecules. 

 

We conclude that, broadly stated, peaceful scientific advances, global statistics and 

demographics of GMOs suggest that the potential for corruption of biotechnology to catastrophic 

malevolent use is considerable. At a more detailed level, we find that there are tangible 

opportunities for many potential adversaries to acquire, modify and then manufactures to scale a 

potential GMO pathogen. Further development of a modified pathogen for use in a full scale 

direct catastrophic biological attack is feasible, but the full spectrum of technologies for scale-up, 

testing, packaging, weapon production and employment will most likely require the resources of 

a nation state or comparably-resourced organization. We recommend that, in concert with 

“science-based “ analysis, further efforts to expand and utilize this analytical framework be 

undertaken to better characterize the future threat from GMOs as well as other emerging threats 

such as those derived from systems or synthetic biology and bioregulators. 

                                                           

1

 We define a catastrophic biowarfare attack as one having direct physical scale, such as the loss of a major U.S. city 

or national system; (As such, the 2001 U.S. Anthrax attacks would not qualify) 
4

CONTENTS 

Introduction ......................................................................................................................................5 

Background ......................................................................................................................................8 

Technical Discussion (Scientific Principles Underlying GMOs) ..................................................14 

   

Framework of Analysis ..................................................................................................................25 

Limited current analysis/Cost-Benefit approach ...........................................................................29 

Preliminary Findings ......................................................................................................................30 

Conclusion .....................................................................................................................................32 

Bibliography 

Annex A. Terms and Definitions 

Annex. B. Recommendations 
5

Introduction 

a. What is the issue? 

Evaluating the potential threat posed by advances in biotechnology, especially genetically 

modified organisms (GMOs), and synthetic biology remains a contentious issue. The rapid 

development of the tools of molecular biology and metabolic engineering has enabled the 

development of chimeric organisms which possess characteristics which are not native to the 

wild variant. This is commonplace in the area of biomanufacturing, where genes are introduced 

into organisms such as E coli and products manufactured via large-scale fermentation. More 

recently, entire metabolic pathways, albeit of limited complexity, have been engineered into 

organisms, for example, for the production of artemisinin in yeast.2 In addition to such metabolic 

engineering projects, whole genomes are being sequenced, leading to the possibility of creating 

organisms de novo. 

Numerous lectures, briefings and articles have argued that the dual use nature of biotechnology, 

the training of foreign students in American universities, and the easy availability of information 

on the internet have given potential adversaries access to biological weapons of unimagined 

which pose an existential threat. Some believe that, inevitably, these advances will lead to a 

catastrophic biological attack. 

Others have argued the opposite that making all information publicly available will enable a 

more universal “white biotechnology” which will ultimately monitor the field and provide the 

means to defeat any threat developed by adversaries. It has been argued that, despite these 

advances, the scientific and technical requirements, as well as the fundamental laws of natural 

selection, will prevent such an attack. 

An example of the controversy is represented by statements such as that found on the web site of 

the Hastings Center, which states that, 

“Research suggests that synthetic biology may soon be a technology of choice for a nation or 

terrorist hoping to develop or acquire a pathogen for use as a weapon”, however, without explicit 

supporting references. 3

To further demonstrate the depth of the issue, a brief listing of the current arguments For/Against 

the likelihood of a catastrophic biological attack being brought about by advances in synthetic 

biology follows. 

Arguments FOR include: 

- Advances in the science and technology of genetics, writ large. 

                                                           

2

 Keasling, Jay D., Production of the anti-malarial drug precursor artemisinic acid in engineered yeast” Nature, 13 

April 2006

3

 Garfinkel, M. et al., http://www.thehastingscenter.org/synthetic-biology-bioethics-briefing-book/ Accessed 30 

August 2010
6

‐ Growth of commercial GMO activities. 

- GMO knowledge base and its availability. 

- Simplicity and availability of required low-cost materials and equipment. 

 - Human abuse of antibiotics and other practices which make populations more 

vulnerable to a GMO. 

- Occurrences of pandemic disease derived from natural genetic evolution. 

Arguments AGAINST include: 

  ‐ Given the complexity of living organisms and their genetic makeup and 

responses, it is extremely difficult to predict the outcome of any genetic modification. 

 - The very limited success of “gene therapy” - peaceful medical objectives of 

genetics for new therapeutics and “individualized” gene based treatments are as yet 

unrealized. 

 - Nature is intolerant of modifications or new organisms and tends to select 

against them. Natural evolutionary processes make/break GMOs continually for the last 

three billion years, and it is unlikely that humans will outdo that. 

- Extreme technical difficulties of “weaponization” for most potential GMO 

pathogens. 

- An unimpressive history of bioterrorist attacks. 

b. Complexity of the task. 

The threat comprises an extremely diverse set of potential actors, tactical and strategic 

objectives, candidate targets to meet those objectives, candidate agent organisms appropriate to 

each, and a wide range of practical approaches for acquiring, modifying, and delivering threat 

organisms to their intended targets. Moreover, the science and technology of GMOs are, and will 

continue to be, a moving target. The field is expanding and knowledge and capabilities 

disseminating globally at a phenomenal rate. Today’s analysis may quickly be overcome by 

other developments in the near future. 

c. Scope of Study and Deliverables.

This analysis focuses on the development of a robust and adaptable analytical framework for 

evaluating the threat posed by genetically-modified microorganisms, particularly those created 

using synthetic biology. The framework also addresses requirements for quantity production, 

packaging and delivery of threat agents to achieve a direct scale of damage against the U.S. at 

the level of biological warfare. 
7

 

The analysis addresses a set of key questions, outlined in subparagraph d. below. Given the 

complexity of the task as described above, we do not present a full answer to these questions. 

Rather, the assessment attempts to lay out the essential technical requirements and alternative 

approaches available for developing a practical threat GMO. These form the point of departure 

for an analytical framework that takes into account the range of potential threat actors and 

objectives. These, together with the analysis of the threat development process, can be used 

develop practical scenarios and evaluate the relative risks and benefits associated with different 

actors and objectives. Such a framework can be used to test various hypotheses and measure our 

depth of understanding. 

d. Key Questions

1. What is the nature and scope of the threat, if any, posed by GMOs, to include the 

potential to develop completely de novo organisms or completely artificial abiotic systems? 

2. What are the fundamental processes and global state of the art for creating GMOs? 

3. Beyond the technical means to create a GMO, what might the follow-on requirements 

for “weaponization” include? 

4. What are the capabilities and incentives for foreign states, transnational groups, small 

terrorist groups, or individuals to attempt to develop a significant GMO threat? 
8

Background

The field of genetic modification of living organisms for human use has undergone explosive 

change. Since its first pragmatic elucidation in 1953, DNA structure and genetic engineering has 

extended its reach into agriculture, animal husbandry, medicine, and even organic materials4

. At 

its scientific limit, researchers are now applying genetic engineering to attempt to create 

completely new or de novo entities outside of the boundaries of normal organic reproduction or 

assembly. This section establishes our view of what a GMO is, provides examples of key and 

recent advances, and then characterizes the size of the effort (market) and its rate of change. 

Multiple references and definitions of GMOs exist. For the purpose of this study, we approach 

the subject in its general form as described below. 

A genetically modified organism is one whose genetic characteristics have been altered by the

insertion of a modified gene or a gene from another organism using the techniques of genetic

engineering.5 Genetically modified organisms encompass a wide spectrum of single and multicellular

organisms, including plants and animals. This effort specifically addresses microorganisms (single cell

biota and viruses). Organisms modified by insertion of genes from another organism are also referred to

as “transgenic” organisms.  

a. Historical and Recent examples of Genetic Engineering.  

While the scope of this assessment focuses on single celled organisms, genetic modification of 

complex multicell organisms is a major commercial activity. To understand the current and 

future path of genetic engineering, it is useful to review some of the initial successes in this field, 

the various areas of application which have ensued, and then, as a subset, several key genetic 

engineering milestones relevant to biodefense. 

Since the early 1990’s genetically engineered plants have been commercially available6

. Socalled “first generation” transgenic plants have been engineered for characteristics that enhance 

the agricultural yield and marketing. Such characteristics include resistance to pests, herbicides 

and extreme climates, as well as improved product shelf life. For example, since their first 

commercial cultivation in 1996, plants have been genetically modified for tolerance to the 

herbicides glufosinate and glyphosate. A “second generation” of transgenic plants, now in 

research and development, is aimed at enhancing consumer satisfaction by enhancing taste, 

                                                           

4

 Nature Archives, A Structure for Deoxyribose Nucleic Acid, Watson J.D. and Crick F.H.C. Nature 171, 737-738 

(1953)

5

 The American Heritage® Medical Dictionary Copyright © 2007, 2004 by Houghton Mifflin Company. Published 

by Houghton Mifflin Company. All rights reserved.

6

 Hails, Rosie S. “Genetically modified plants – the debate continues”, Institute of Virology and Environmental 

Microbiology, Oxford, UK; Tree Volume 15, 1 January 2000
9

texture, or appearance of produce. To date, no second generation transgenic plants are on the 

market. 

Genetically modified/transgenic animals are used in a wide range of applications. Simple 

organisms such as fruit flies have been used to study the effects of genetic changes across 

generations. Transgenic mice are often used to study cellular and tissue-specific responses to 

disease. 

Transgenic bovines and goats have also been developed to express a variety of useful 

biologically derived products. Among the first of these was “Herman the Bull”, who was 

genetically modified in 1990 with a human gene sequence while in embryonic form to produce 

lactoferrin, an immune system protein7

. This was followed by the development of a transgenic 

goat that expressed proteins for silk (similar to spider silk) developed by the Canadian firm, 

Nexia, under the trade name BioSteel TM.

8

 On February 6, 2009 the U.S. Food and Drug 

Administration approved the first human biological drug, also extracted from goat’s milk. The 

drug, ATryn, is an anticoagulant which reduces the probability of blood clots during surgery or 

childbirth. 9

 Gene therapy, involving the use of viruses as a vector for introducing generic material 

into cells, has had some success in treating genetic disorders such as severe combined 

immunodeficiency, and treatments are being developed for a range of other currently incurable 

diseases such as cystic fibrosis, sickle cell anemia, and muscular dystrophy. Genes introduced in 

this manner are not transmitted to the next generation. Gene therapy targeting the reproductive 

cells—so-called “Germ line Gene Therapy”—at present carries an unquantifiable risk associated 

with interfering with other genes, hence near-term development and commercialization of this 

technology is unlikely.10 

 In 2009, scientists in Japan announced that they had successfully transferred a gene into a 

primate species and produced a stable line of breeding transgenic primates for the first time. 11

Genetically modified bacteria have become commonly used as a means for producing large 

amounts of pure human proteins for use in medicine. Examples include production of insulin to 

treat diabetes, clotting factors to treat haemophilia, and human growth hormone to treat various 

forms of dwarfism. In addition, advances in biomedical research continue to build a growing 

store of knowledge directly applicable to the development of GMO threat agents. In recent years 

there have been a number of substantial development bearing on the potential threat of 

                                                           

7

 M.F. Brink, Developing efficient strategies for the generation of transgenic cattle which produce 

biopharmaceuticals in milk”, Theriogenology, An International Journal of animal Reproduction, Volume 53, Issue 1 

Pages 139-148, January 2000

8

 Vendrely, Charlotte, Biotechnological Production of Spider-Silk Proteins Enables New Applications, 

Macromolecular Biosciences, volume 7, Issue 4 pages 401-409. April 10, 2007

9

 Erickson, Britt (10 February 2009). FDA Approves Drug From Transgenic Goat Milk. 10 American Journal of Law and Medicine, FDA Regulation –An Answer to the Questions of Human Cloning and 

Germline Gene Therapy, Boston University School of Law, 2001 11 Cyranowski, David, Marmoset Model takes Centre Stage, Nature, 459-523-527, May 2009
10

genetically modified or synthetically produced microorganisms. Genetic research in the field of 

biodefense relevant activities has also flourished. Several key examples follow. 

• In 1981 scientists cloned a full-length virus genome (Poliovirus) that was infectious to 

mammalian cells and demonstrated the basis of the ability to replicate an infectious RNA 

virus.12 Significantly, in 2002, Cello et al. reported purely chemical synthesis of an 

infectious Poliovirus in the absence of any natural template.13 

• The potential for modifying organisms to significantly enhance virulence and mortality 

rate was shown when modified mousepox virus [in the same family (Poxviridae) as 

smallpox], intended for use as a contraceptive, proved 100% deadly by circumventing 

host immune defenses, even in previously immunized (vaccinated) animals.14 

• Genetic information even for highly virulent pathogens is widely available. The complete 

genome sequence of 45 variola strains providing supplemental material with gene 

organization of smallpox is freely available on the web.15

• Cloning and recovery of infectious Ebola virus and of a mutant more cytotoxic than the 

natural wild-type.16

• Later generation of a complete infectious genome (5,400 bases long in a bacteriophage) 

from synthetic oligonucleotides synthesized according only to the sequence reported in 

GenBank. The synthesis and assembly of this organism were completed in only 14 days 

and without a need for accessing any living organism.17 

• In 2008, Israeli researchers published a procedure for the de novo construction of errorfree DNA molecules from error-prone commercially available oligonucleotides.18 This 

ability was cited as having the potential to allow masking of an intended synthetic 

molecule or organism during purchase of oligonucleotides. 

• In 2010, the J. Craig Venter Institute reported the successful synthesis of a complete 

microbe genome comprising over 1.0 million base pairs, and insertion of same into a 

microorganism capable of reproducing. However, assertions that this constitutes a fullysynthetic life form are arguably overstated, but the demonstrated ability to replicate the 

                                                           

12 V.R. Racaniello, D. Baltimore, Cloned Poliovirus complementary DNA is infectious in mammalian cells. Science, 

1981, new Series (4523), 916-919

13 Paul Cello, E Wimmer, Chemical synthesis of poliovirus cDNA: Generation of infectious virus in the absence of 

natural template. Science, 2002, 9 (297), No. 5583 14 R.J. Jackson, et al. Expression of mouse interleukin-4 by recombinant ectromelia virus suppresses cytolytic 

lymphocyte responses and overcomes genetic resistance to mousepox. Journal of Virology. 2001, 75 (3) 15 J.J. Esposito et al., Genome sequence diversity and clues to the evolution of variola (smallpox) virus. Science, 

2006, 313, 807-812

16 V. E. Volchkov et al., Recovery of infectious Ebola virus from complementary DN: RNA editing of the GP gene 

and viral cytotoxicity., Science, 2001, 291, 1965-1969. 17 H.O. Smith et al., Bacteriophage from synthetic oligonucleotides., Proc.Natl.Acad. Sci. U.S.A. 2003, 100 (26), 

15440-15445.

18 G. Linshiz et al., Recursive construction of perfect DNA molecules from imperfect oligonucleotides. Molecular 

systems biology 4: 191; doi:10.1038/msb.2008.26]
11

full DNA sequence is a substantial accomplishment. Hence, the claim would be better 

qualified as the assembly of a very simple life form by removal and addition of already 

living materials into a single cell structure. 

b. How large is the GMO market and global enterprise? 

Since the initial elucidation of the structure of DNA in 1953, and despite some market resistance 

(non-GMO attitudes, government and academic restrictions) the market for GMO products has 

rapidly expanded. Agricultural, chemical, pharmaceutical, industrial biotechnology (including 

energy) fields have all benefited from GMO activities and experienced levels of growth that 

outpaced other national employment.19 In 2009, commercial growth in the U.S. produced income 

of $75 billion dollars to a group of approximately 650 bioscience companies.20

 As an example, the global commercial value of biotechnology crops grown in 2008 was 

estimated to be $130 billion. 21 The United States Department of Agriculture (USDA) reports on 

the total area of GMO varieties planted. According to the National Agricultural Statistics 

Service, the states published in these tables represent 81–86 percent of all corn planted area, 88–

90 percent of all soybean planted area, and 81–93 percent of all upland cotton planted area 

(depending on the year).22

Underlying this commercial growth are significant advances in the state of the art in genetic 

engineering and synthetic biology. The field has benefited from the confluence of several 

important technological trends: The discovery and subsequent rapid development of the field of 

genomics, the development of new tools and techniques for inspection and manipulation of 

matter at molecular levels, coupled with advances in information technology enabling efficient 

storage, processing, and dissemination of the vast amounts of data generated by advancing 

research in these areas. 

 The Number of DNA “synthesis foundries” world-wide has continued to grow. As of 

November 2010 there are an estimated thirty countries capable of synthesizing genes or genetic 

sequences of 1000 base pairs or larger. Including private, government and academic gene 

synthesis organizations a partial open source list yields: China (36), Germany (20), Great Britain 

(14), France (9), Russia (8), India (6), Canada (7), Netherlands (6), Israel (1), Iran, (1), and 

several others.23

 Private DNA “synthesis foundry” companies’ operations and reach are multinational. 

Eurofins MWG Operon has foundries in the US, Germany, and India; Takara Biotechnology 

(Dalian) in China is a subsidiary of Takara Bio Inc. of Japan; ThermoFischer of the US has a 

foundry in Lithuania; Origene operates foundries in both the US and China; and the Zelinsky 

                                                           

19 U.S. Biosciences employment growth 2001-2008, from 100K to 600K Battelle, 2010

20 Battelle, 2010 Biotechnology Summary; Net Income 2009; dated 2010

21 Clives, James. Global Status of Commercialized Biotech/GM Crops 2008, ISAAA Brief 39 22 USDA Economic Research Service, Adoption of Genetically Engineered Crops in the USA, Table 

http://www.ers.usda.gov/Data/BiotechCrops/

23 Defense Intelligence Agency, Unclassified Memorandum – NCI Response to ECBC, 9 November 2011
12

Institute Inc. of the US markets the products and capabilities of the Zelinsky Institute of Organic 

Chemistry in Russia. 

The breadth of examples of key scientific breakthroughs described earlier offer evidence of 

scientific to commercial strength and public acceptance, and suggests that biological engineering 

is coming to be viewed in a manner similar to that of traditional engineering. The technical 

events and markers in the area of microorganisms demonstrate the practicality of manipulating of 

microorganisms and pathogens to change their characteristics. The advent of synthetic biology 

holds promise for novel and perhaps completely artificial, “abiotic”, functioning cells. 

c. Rate of Change for Biotechnology/GMO technology 

The rate of change for Biotechnology/GMO technology can be appreciated by considering job 

growth and the economics of GMO technologies. With respect to job growth in the U.S., 

between 2001 to 2008 employment in the bioscience fields rose from 100,000 jobs to over 

600,000 (600%).24 Considering the economics of GMO technologies, broad and growing markets 

for all types of GMOs, coupled with the widespread availability of basic information and 

technology, have driven rapid development and dissemination of the technology. The costs of 

both sequencing and synthesizing genetic material (key enabling capabilities for GMO 

development) have dropped dramatically in recent years (See figure 1.) and these trends are 

expected to continue. At present, costs for synthesis of short sequences of DNA are running as 

low as 0.3 Euros (40 cents/base pair), and costs for sequencing in 2010 are approaching 

$1.0/million base pairs. 25 As a point of reference, there are some 2.9 billion base pairs in the 

haploid (chromosomal) human genome. 

Figure 1. “Carlson Curve”. http://www.synthesis.cc/

These include modifications based entirely on synthesized DNA where, as outlined above, 

the sequence length of purely artificial DNA has grown from a few thousand base pairs to 

over 1.0 million in the past decade. 

                                                           

24 U.S. Biosciences employment growth 2001-2008, from 100K to 600K Battelle, 2010

25 Carlson, Robert: as quoted in the On-line Economist, Date 12 August 2010. 
13

This significant rate of change reflects the growing social awareness of global health issues and 

the increasing footprint of the pharmaceutical, medical, agricultural and industrial biotechnology 

sectors in national economies (not necessarily related). This has resulted in both rapid growth 

and globalization of genetic engineering capabilities. GMO advances to identify, characterize, 

modify, fabricate (clone, in-vitro replication, etc.), and stabilize DNA have all advanced rapidly 

and substantively. In this context, many of the arguments in favor of GMO threat expansion are 

understandable. 
14

Technical Discussion: Scientific Principles Underlying GMOs

This section details the microorganisms of interest in this study, and then outlines the basics of 

genetic modification of microorganisms and the availability of various means for developing 

potential threat agents. 

a. Microorganisms of interest 

1. Bacteria. Bacteria are unicellular organisms. The unit of life is the cell, the smallest entity 

capable of displaying attributes associated with the living state, including growth, metabolism, 

stimulus response and replication. Bacteria can be modified by conducting modification of the 

cell while in a host organism, altered by chemical and/or organic means, or modified by plasmid 

introduction (circular DNA without DNA synthesis). Of the options available, plasmid 

introduction provides the most predictable product and outcomes and, by comparison, is 

inexpensive and technically trivial. However, stability of the plasmids over generations is not 

ensured. 

2. Viruses. Although viruses are not technically alive, they represent supramolecular assemblies 

that act as parasites within host cells, underscoring the functional “culling out” of specific 

cellular processes, albeit within the confines of living cells. Viruses can be modified via DNA or 

RNA manipulation, where virulence factors can be spliced into the virus. Other modifications 

include changing the viral protein coat such that they can target specific type cells. 

3. Listed but not included in Framework Analysis 

 * Multi-cell pathogens 

 * Toxins (Chemical products of living cells.) 

 * Fungi (Robust organism; no genetic manipulation needed) 

 * Prions ( Generally not subject to genetic modification) 

b. Options for acquiring pathogenic microorganisms for use as biological agents 

Acquisition and use of naturally-occurring pathogens is a viable baseline for the development of 

a rudimentary biological agent. An example of a naturally-occurring organism that has been 

promoted as a biological agent both by nation states and terrorists is anthrax. The primary 

challenge is that acquisition of cultures “in-the-wild” requires visiting an area where the disease 

of interest is active. However, given an opportunity, the necessary samples can be collected and 

preserved by any adequately trained technician. Alternative methods, in general ascending order 

of difficulty include: 

 * Obtaining an agent from a research center where work is being performed. 
15

 * Ordering from one or more culture collections maintained world-wide. 

 * Creating it either by modifying another pathogen or, in the case of viruses, synthesizing 

it from its obtainable components using conventional gene-splicing techniques or outsourcing to 

a DNA sequencing companies (also referred to as DNA foundries). 

 * As noted in the Background, RNA and DNA sequences for both viruses and 

microorganisms have been synthesized entirely from their genetic code. 

 * Finally, synthetic biology, including ongoing research in so-called “protocells” 

The emergence of DNA foundries adds a new dimension to the potential threat. In the past, 

research activities have extracted and used gene splicing techniques to modify genetic materials, 

an approach which is labor intensive. DNA/Gene synthesis is being widely advertised as a more 

cost effective, less time-consuming approach. 

Culture collections with pathogen stocks exist in many countries. General DNA materials 

(including possible pathogen stock) are normally developed or established as a unit “culture 

stock”, not as a single cell or bacterium. Culture stocks are, by standard, a test tube sized colony 

of the pathogen in a slant auger gel. The culture stock must be maintained at – 80 to -90 degrees 

F until it is needed for amplification, at which time it is thawed and grown in an appropriate 

medium, such as a 25-50 liter fermentation tank. 

The World Federation for Culture Collections, World Data Center for Microorganisms database 

lists some 581 Culture Collections in 68 countries, holding over 1.6 million culture samples. In 

descending order, based on number of culture centers, the countries listed are Brazil (60 centers) 

, Thailand, France, Australia, Japan, India, China, USA, Canada, the UK, Indonesia, Mexico, 

Russian Federation, and Republic of Korea (15). In terms of number of cultures maintained, the 

leaders (again in descending order) are: The US (210,276), Brazil, Japan, Denmark, United 

Kingdom, Netherlands, Australia, China, Republic of Korea, Canada, France, India, Belgium, 

Sweden, Germany, and the Russian Federation (45,655). 

Most major academic institutions and national governments have some level of involvement in 

research. The Militarily Critical Technologies Program (MCTP) has estimated that there are over 

400 locations around the world that maintain cell cultures that might be used as starting points 

for biological agent development, and biological materials from these culture collections are 

generally available to research centers world-wide. A significant consideration in terms of 

government oversight and security is fewer than half of the 581 collections are in government 

facilities. 26 

The statistical breakout is shown in Figure 2. 

                                                           

26 WDCM Statistics Web Site, http://wdcm.nig.ac.jp/statistics.html, dated 17 December 2010. 
16

Number of Cultures Held World-Wide

Bacteria

46%

Fungi

31%

Other 

microbials

21%

Cell Lines

1%

Viruses

1%

Bacteria

Fungi

Viruses

Cell Lines

Other microbials

 Figure 2. Number of Cultures Held World-Wide 

 

c. Methods to modify or create a threat GMO

Synthetic biology holds potential for engineering pathogenic organisms not found in nature, 

either by introduction of synthetic genes into an existing natural organism, or by completely 

artificial “abiotic” synthesis of organisms. The potential effectiveness of microorganisms as 

biological agents can be enhanced in a number of diverse ways: 

 * Modify the organism by putting a molecule on the coat of an existing pathogen that 

causes it to bind more efficiently to the host target cell. 

 * Insert genetic material that encodes for a toxin (typically a protein). An example is the 

insertion of a plasmid with the 0157H gene in E. Coli. The genome of the cell isn’t altered, but it 

carries genetic material in the plasmids that encode the expression of toxins. 

 * Modify the genome of the microorganism itself to increase infectivity and virulence. 

 * Insert DNA sequences that bind to the host so that the host’s immune reaction is 

suppressed. 

 * Create a completely artificial or “abiotic” pathogen. Unlike current synthetic biological 

approaches, an abiotic organotypic approach would abstract the functionalities of living systems 

without copying their components.27

                                                           

27 Dr. James Valdes, Ph.D., “Transformational Countermeasures Technology Initiative”, RDECOM, 2008
17

 

The genetic material for modification may be either derived from natural organisms using 

standard recombinant DNA techniques, or produced by DNA synthesis, the latter being much 

less labor intensive. In recent work, DNA sequences on the order of 1 million base-pairs have 

been synthesized entirely from digitized genome sequence information, and the resulting 

organisms were phenotypical and capable of self-replication.28 

As is discussed in greater detail throughout this report, the tools and information required for 

genetic modification of microorganisms are readily available worldwide. The growth of synthetic 

biology, with its “engineering approach”, is expected to eventually lead to development of cheap, 

highly standardized building blocks (e.g., so-called Bio-BricksTM) and the design rules required 

for their functional assembly being widely disseminated. 

d. Personnel and Costs

Assertions to the effect that a high school graduate can develop an effective biological weapon 

are arguably overstated. However, the work can be successfully accomplished by a small cadre; a 

trained clinician (if the feedstock is gathered in the wild), a graduate microbiologist, and a good 

laboratory technician are probably an adequate minimum staff. Depending on the nature of the 

intended attack, quantities of modified natural organisms sufficient to infect an individual, a 

small area or small concentrated group are within the capability of an appropriately trained 

individual, such as a competent microbiologist or medical clinician. 

Development of novel (i.e., not known to be naturally-occurring) GMOs exhibiting unique 

designer characteristics requires substantially greater knowledge and capability. Many 

industrialized nations have laboratories capable of analyzing which immune response modifier 

genes in humans and livestock, when inserted into an organism together with pathogenicity (e.g., 

adherence and invasive) factor, will yield highly infectious pathogenic organisms. 

In the development of biodefensive measures, however, the cost associated with characterizing 

genes of various species, including humans, that increase disease resistance or susceptibility is 

relatively high. There are approximately 25,000 genes in the human genome. The determination 

of the most preferred assembly of genes that will yield very high or very low susceptibility to 

infection will require significant financial commitments and fairly sophisticated research skills. 

Offsetting the cost, results of research in this area is widely published and the resulting 

information stored in open-source repositories. The bioinformatic tools and techniques to access 

and manipulate the data are likewise widely shared globally. 

e. Skills required to modify or create a threat GMO

The skills required to use a GMO as a threat falls into four broad categories. The first three, gene 

mapping, functional genomics, and bioinformatics, broadly comprise the discipline of genomics. 

The fourth category is, at the current state-of-the-art, less well developed. It comprises a loosely-

                                                           

28 Gibson, D. G., et ales; Creation of a bacterial cell controlled by a chemically synthesized genome. Science, 2010 

Jul 2; 329(5987):52-6. Epub 2010 May 20. 
18

defined set of subspecialties such as pharmacogenomics, toxicology, immunology, biostatistics, 

epidemiology, and biochemisty. The common thread of these is the central role of understanding 

how organisms react to infection and to exposure to toxins, and broadly define the area now 

known as systems biology. 

Information, particularly regarding naturally-occurring organisms and gene sequences, is freely 

accessible. As noted in previous studies29 the genetic information on a vast number of 

microorganisms as well as animals, plants, and humans is well structured, largely standardized, 

and accessible at www.ncbi.nih.gov/Genbank/ and other bioinformatics repositories. GenBank, 

with the DNA Data Bank of Japan, and the European Nucleotide Archive (formerly the 

Eurpoean Molecular Biology Laboratory Nucleotide Sequence Database) participates in the 

International Nucleotide Sequence Database Collaboration, an cooperative effort to gather and 

disseminate nucleotide sequence and annotation and links for the three major data repositories. 

The INSD Collaboration has a uniform policy of free and unrestricted access to all of the data 

records their databases contain. Scientists worldwide can access these records to plan 

experiments or publish any analysis or critique. No use restrictions or licensing requirements are 

included in any sequence data records, and no restrictions or licensing fees may be placed on the 

redistribution or use of the database by any party. This means that the sequences of many 

potential threat biological agents are freely available, and the information needed to synthesize or 

modify them is freely accessible. Anyone may access the computational tools to design 

genetically engineered organisms free of charge. 

The latest update to the Biomedical Section of the Militarily Critical Technologies List 30 also 

identifies the following countries as having large scale datasets related to genetic engineering: 

Australia, Germany, Italy, Japan, The Netherlands, Sweden, and the United Kingdom. 

As of April 2010, the International Nucleotide Sequence Database repository of DNA sequences 

exceeded 100 gigabases.31 In comparison, the whole genome of Ebola virus is approximately 

19,000 bases. The Web also provides free access to bioinformatics applications and software 

tools for analyzing genomic information. There is also a growing body of knowledge and data on 

proteomics, thus linking nucleic acid sequence information with the biological functions of 

proteins. These databases and tools characterize the function of specific genes with everincreasing detail and fidelity, coupled with the ability to “mail order” sequences from a growing 

number of DNA sequencing companies world-wide provide a baseline capability for genetic 

engineering of microorganisms. 

For example, the European Bioinformatics Institute [www.ebi.ac.uk] (EBI), part of the European 

Molecular Biology Laboratory (www.embl.org/), provides on-line access to a comprehensive 

range of tools for the field of bioinformatics (over 135 are currently listed). Online information 

available includes: 

                                                           

29 Sagripanti, Jose-Luis, Ramsbotham, Alan J. ECBC – TR-666 – “Global Survey of Research and Capabilities in 

Genetically Engineered Organisms that could be used in Biological Warfare or Bioterrorism” Edgewood Chemical 

and Biological Center, December 2008.

30 Militarily Critical Technologies List, Section 4, Biomedical Technology, June 2009

31 http://www.insdc.org/documents/feature_table.html
19

 

 * Similarity and Homology - the BLAST or Fasta programs can be used to look for 

sequence similarity and infer homology. 

* Protein Functional Analysis - InterProScan can be used to search for motifs in a protein 

sequence of interest. 

 * Proteomic Services NEW - UniProt DAS server allows researchers to show their 

research results in the context of UniProtKB/Swiss-Prot annotation. 

* Sequence Analysis - ClustalW a sequence alignment tool.

* Structural Analysis - MSDfold or DALI can be used to query any protein structure and 

compare it to those in the Protein Data Bank (PDB). 

* Web Services - provide programmatic access to the various databases and 

retrieval/analysis services. 

* Tools Miscellaneous - Expression Profiler a set of tools for clustering, analyses, and 

visualization of gene expression and other genomic data. 

Answers to most technical questions that may arise can be found on the World Wide Web at one 

of the help sites from the many universities that carry related activities within newly formed 

departments of bioinformatics or specific informatics resources. As the global knowledge base 

characterizing functional genomics and proteomics expands and is dispersed to less developed 

nations, it will be relatively easier and inexpensive to generate genetic combinations that will 

markedly increase infectivity and pathogenicity of any given organism. 32 

f. Likely technical objectives of modifying or creating a threat GMO. There are a number of 

objectives, each with their own technical requirements and technical difficulties that a 

perpetrator might pursue. These include, but are not limited to, either uniquely or in 

combination: 

 * Increase infectivity. 

 * Increase virulence/mortality rate. 

 * Diminish host immunity or confer antibiotic resistance. 

 * Increase survivability outside the host (i.e., environmental stability). 

 * Circumvent/degrade detection/protection measures. 

A key observation however, is that the first several bullets drive a trade-off with regard to the 

quantity of material needed to pose a threat. Threats with a very high virulence and mortality 

                                                           

32 MCTL Section 4.1 Host Genome Material in Virus-like Agent to Affect Soldier Capability, June 2009
20

rate, while psychologically devastating, tend to be self- limiting unless they also have high 

person-to-person infection rates, in which case they may take a while to “burn out.” In practice, 

the objectives of genetically-modifying organisms for use as biological agents will be driven by 

the user’s perceived threat/benefit analysis, difficulty of production, and the threat characteristics 

that they possess. 

 

 g. Catastrophic Attack and Weaponization

 1. Weaponization involves co-opting the advances of genomics for malevolent purposes. 

For the objectives of this study, deployment of such weapons might result in catastrophic 

consequences. Further, we define a catastrophic attack on the basis that there is a direct scale 

consequence of the attack. This is unlike a typical terrorist attack where the main effect is 

psychological. For example, we do not consider the 2001 U.S. Anthrax Attacks to be 

“catastrophic”. Although there were significant indirect consequences, the actual numbers of 

people killed were very few. We would define a significant direct attack as one which results in 

thousands of casualties up to the loss of population equivalent to that of a major U.S. city. 

Weaponization is the most difficult task in conducting biowarfare. Although it may be possible 

to create or modify a pathogen in a laboratory, the next steps of producing sufficient quantities of 

the pathogen, deriving a means to take it out of the lab and have it survive to the point of attack, 

and then to disseminate it successfully all pose significant challenges. Within those steps there is 

requirement for a means of delivery which is timely, sufficiently broad and with an effective 

“uptake” or infectivity in transmission. Weaponization techniques can vary greatly. As examples, 

one threat pathogen may require scale production, stabilization via inert coating for 

“encapsulation” and then aerosol distribution via mechanical means; another less controlled 

approach would be through contagious vectors, infecting and then releasing carriers of the 

pathogen to naturally replicate and deliver the threat. This section covers principles which will 

be further discussed in the next section on Framework of Analysis. 

2. What does it take to produce a volume of pathogens to the scale needed for a 

significant or catastrophic attack? Depending on the type of pathogen, the amount needed for a 

catastrophic attack could range from several milliliters to 55 gallon drums. The respiratory doses 

for various microorganisms for an infection in humans (measured in ugs) range from 0.00000021 

for Q-fever to .008 for anthrax. Respiratory doses for biotoxins such as Staphylococcal 

Enterotoxin B (SEB) (0.025 ugs) and Botulinum neurotoxic (4.5 ugs) are orders of magnitude 

higher. For comparison, the effective dose for the Nerve Agent VX is 70ug; many orders of 

magnitude greater than that of Q-Fever. 33 

3. Materials. The materials required such as reagents, culture media and host vectors are readily 

available worldwide. These are all used in a variety of life sciences and environmental 

applications, and there are no effective restrictions on a potential enemy’s access to them. 

                                                           

33 Dr. Robert Armstrong, Dr. David Franz, Dr. James Valdes, Bill Patrick’s Relative Aerosol Potency Chart, 

Biological Agents: Threat, Preparedness, Response and Myths, presentation to European Commission, February 

2009
21

4. Facilities. The cost of a facility for modifying, culturing, and replicating GMOs in quantity 

sufficient to pose a significant biological agent threat has been estimated by independent studies 

to be on the order of $200K to $25OK. With such a facility and using proper scale-up 

bioprocessing techniques, one can amplify the volume of a test tube culture sample to a 25 – 50 

liter basis within 24 hours.34

 Table 1. Money * Genetic Engineering Comparative Costs 

Compared to other projects that might be undertaken by governments or private organizations, 

the cost of equipping and staffing a laboratory scale bioprocessing facility, as shown in Table 2 

below are trivial. 

Table 2. Comparative Costs for a scale bioprocessing facility 

                                                           

34 The data and discussion of Tables 1 and 2 are extracts from Sagripanti, J-L, et al. Global Survey of Research 

Capabilities in Genetically Engineered Organisms that Could be Used in Biological Warfare or Bioterrorism., ECBC 

Technical Report ECBC TR-666. December 2008. 
22

5. Ability to stabilize pathogen for delivery. Assuming an adversary is capable of 

accomplishing genetic modification and scale production of a threat pathogen, outside of an 

infectious disease model, the agent must then be prepared to be used as a weapon. Given the 

ambient conditions of sunlight, temperature, and exposure to other meteorological factors, most 

microorganisms do not survive unless specially prepared. The next critical step is therefore to 

stabilize the pathogen in a form that allows for such survival. Previous techniques included 

“microencapsulation”, in which the pathogen is coated with a protective material as, for example, 

in the coating of enzymes for laundry detergent; embedding in biofilms; and, in rare cases, use of 

living vectors in a manner similar to, but more controllable than, an infectious disease model. 

 6. Means of Delivery. Finally, to launch a catastrophic attack, the perpetrator must have 

some means of delivery. Possible methods include direct means of delivery via aerosol, indirect 

means via packaging and leveraging of US delivery systems (e.g., FEDEX, US grocery 

distribution system, Postal Service) and delivery via natural vectors. 

h. Effectiveness of the tools of synthetic biology for threat GMOs. 

As one respected scientist summarized: “Today, anyone with a high school education can use 

widely available protocols and prepackaged kits to modify the sequence of a gene or replace 

genes within a microorganism; one can also purchase small, disposable, self-contained 

bioreactors for propagating viruses and microorganisms. Such advances continue to lower the 

barriers to biologic-weapons development.35“ 

Is this really true and, if so, how far is the barrier to biological weapons development being 

lowered? Being able to modify genetic material is one thing; understanding the end effects in 

terms of how such modification will affect the characteristics of the organism and its effects on a 

host organism’s physiology is something very different. There is strong evidence to suggest that 

intentional modification of a pathogen remains difficult. 

From the point of view of the potential perpetrator, the challenge is to reliably predict the overall 

effects of the changes. For example, the intent of the Australian researchers in modifying mouse 

pox was to produce a contraceptive effect, and the subsequent lethality of the modified virus was 

an unintended side effect. Conversely a sequence of genetic material that codes for expression of 

a particular toxic protein may inadvertently suppress other functions essential to the reproduction 

or survivability of the microorganisms. 

Arguments against GMO weapons include the limited success of “gene therapy”. During its 

inception, pursuant to the derivation of the human genome and advanced bioinformatics and 

DNA processing, it was posited that unique individual genomes would be determined and then 

used to prescribe gene-based therapeutics for a variety of diseases. With a few general 

exceptions, individualized gene therapies have not yet emerged. This lack of success is generally 

attributed to the observation that human genomics does not imply a specific one-to-one mapping 

of particular genes to singular specific health responses. Instead, the human gene composition 

                                                           

35 David A. Relman, “Bioterrorism – Preparing to Fight the Next War,” The New England Journal of Medicine 354, 

no. 2 (2006) 
23

includes numerous redundancies where multiple genes or a system of backup genes can all play a 

role in immunity and response to a pathogenic challenge. Organisms and their genetic 

composition and host-pathogen interactions are exceedingly complex. 

As a further example, analysis of one of the simplest pathogens, the prion, was conducted 

utilizing the latest methods of systems biology. Using multiple mouse models, gene expression 

data, and techniques such as subtractive biology, an initial set of more than 7,400 genes whose 

expression changed in response to prion infection was winnowed down to 333 which were 

critically involved in disease progression, and specific multiple effects on metabolic pathways 

were determined. 36 Such a systems biology approach could eventually lead to very targeted 

medical countermeasures, either prophylactic or therapeutic, but could also be used to predict or 

target the effects of a pathogen. 

With respect to the de novo design of entirely new GMOs, there are additional challenges based 

on the spatial architecture and geometry of the cellular environment.37 In this case, the intricacies 

of having cellular structures and processes come together in the correct spatial/temporal points to 

achieve proper function is an exceptionally difficult challenge. Building a synthetic cell, or even 

making a drastic modification to an existing cell, must account for this architecture. 

Finally, to date, the results of previous biological attacks have been most unimpressive. The most 

recent instance was the 2001 anthrax mailings where, despite the perpetrator using a weapons 

grade pathogen and using the U.S. mail system for physical delivery of the anthrax spores, the 

results of over one billion doses mailed was five deaths.38 

In summary, developing a pathogen suitable for use as a biological weapon agent and its 

subsequent “weaponization” is not simple. Acquisition or creation of a pathogen, subsequent 

genetic modifications, amplification of the pathogen stock to a volume that is subsequently 

stabilized and delivered in a naturally hostile environment against a well-defended public 

accumulates many challenges. 

                                                           

36 Leroy Hood, Finding Early Signs of Mad Cow Disease, Molecular Systems Biology, March 2009 37 Ochman and Raghavan, “Excavating the Functional Landscape of Bacterial Cells; Science 27 November 2009 38 The Unimpressive History of Bioterrorist Attacks, slide 21 Biological Agents: Threat Preparedness, Response and 

Myths
24

Framework of Analysis

a. Why a framework and for what is it used?

Why? Given the complexity and uncertainty of this issue, one cannot leap to a single, or even a 

multiple set of answers. A framework serves as a functional alternative to structure our thinking 

and understanding of the issues and ultimately serves as a platform from which to develop 

particular answers. The framework reflects the key dimensions of consideration which are used 

to “hang” ideas or facts on, test various hypotheses and measure the depth of our understanding. 

What? Framework consists of key questions, subordinate metrics and facts needed to answer 

those questions. 

The framework emphasis is on single cell organisms, viruses, and a limited group of multicellular fungal organisms that pose potential threats (such as those causing plant rust.) The 

framework developed includes placeholders for future elaboration on the threats posed by such 

organisms to multi-cellular life forms. As our understanding of the functioning of different 

metabolic pathways and the damaging effects of their products (such as proteins) improves, these 

placeholders can be expanded to assess whether the increased utility of GMOs relative to 

naturally-occurring organisms is likely to provide an incentive for their development. 

b. Framework structure.  

For the purpose of this study, our Analytical Framework is predicated by the current set of 

Technical Facts and Requirements to create and expand GMOs from single culture to scale 

distribution. Using that set of preliminary facts we then constructed a two dimensional 

framework to reflect the process chain necessary to conduct a deliberate scale bio-attack with a 

GMO, and then to assess the more subjective elements of “outcomes” as a function of 

“feasibility effects” and “cost-benefit” analysis The technical predicate and framework are 

depicted in the following graphics: 

Technical Facts / Requirements to

create or modify GMOs from

single culture to scale

distribution
25

   

At the “meta level” these include 6 parametric variables: 

 1. Who are the likely perpetrators? 

 2. What it takes to make a single GMO culture stock? 

 3. What it takes to make scale quantities for a catastrophic attack? 

 What is takes to “weaponize” a GMO? 

 4. Embed pathogen 

 5. Means of delivery / dissemination 

 6. What are the potential GMO outcomes (Feasible Effect & cost/Benefit)? 

Beneath each of these variables are additional subordinate variables, further amplifying the 

possible permutations and combinations of approaches and outcomes. 

1. Who are the likely perpetrators? 

 * “Grad student” accident or experiment 

 * Lone terrorist 

 * Disgruntled employee 

Perpetrator

‐ Student

‐ Lone Terrorist

‐ Federal Employee

‐ Small Group

‐Transnational

Terrorists

‐ National Opponent

‐ Coalition of enemies

‐ “Mother Nature”

GMO Threat Analytical Framework

One Culture Deliver Scale‐up Embed

Weaponize

Feasible

Effect

Cost /

Benefit

Acquire

Pathogen Stock

• “wild type”

• research

center

• DNA Foundry

• Lab synthesis

• “de novo”

Modify for/to

pathogen

• Bacterium

• Virus

• Toxin

• Fungus*

• Prion*

Modify to avoid

Detection

Determine

Toxicity for

desired effect:

Bacterium

• Culture

medium

• Fermenter

Virus

• ??

•

Toxin

Fungus

Prion

•“In vivo”

human

•Vectors

• Encapsulate

• Corrupt

medical supply

• “Reservoirs”

•Infection

• Dispersion

‐ Aerosol

‐ Fedex

‐ Grocery

logistics

• Medical

logistics

• Vectors

• Poisoning

• Cross‐

contaminatio

n

• Human

Casualties

• Health Care

System

• Agriculture /

Animals

• Area Denial

• Equipment

Contamination

• Psychological

• Economic

Technical Process Chain

• Cost overall

• Probability

of effect

• Relative to

non‐GMO

effects?

• Safety of

perpetrator

• Forensic

signature;

• Possibility of

retribution

Outcomes

Perpetrator

‐ Student

‐ Lone Terrorist

‐ Federal Employee

‐ Small Group

‐Transnational

Terrorists

‐ National Opponent

‐ Coalition of enemies

‐ “Mother Nature”

GMO Threat Analytical Framework

One Culture Deliver Scale‐up Embed

Weaponize

Feasible

Effect

Cost /

Benefit

Acquire

Pathogen Stock

• “wild type”

• research

center

• DNA Foundry

• Lab synthesis

• “de novo”

Modify for/to

pathogen

• Bacterium

• Virus

• Toxin

• Fungus*

• Prion*

Modify to avoid

Detection

Determine

Toxicity for

desired effect:

Bacterium

• Culture

medium

• Fermenter

Virus

• ??

•

Toxin

Fungus

Prion

•“In vivo”

human

•Vectors

• Encapsulate

• Corrupt

medical supply

• “Reservoirs”

•Infection

• Dispersion

‐ Aerosol

‐ Fedex

‐ Grocery

logistics

• Medical

logistics

• Vectors

• Poisoning

• Cross‐

contaminatio

n

• Human

Casualties

• Health Care

System

• Agriculture /

Animals

• Area Denial

• Equipment

Contamination

• Psychological

• Economic

Technical Process Chain

• Cost overall

• Probability

of effect

• Relative to

non‐GMO

effects?

• Safety of

perpetrator

• Forensic

signature;

• Possibility of

retribution

Outcomes
26

 * Small group (need not be transnational or have affiliation with any recognized 

group) 

 * Transnational terrorist or adversarial group 

 * National opponent 

   * Coalition of enemies 

 * Mother Nature 

2. What does it take to acquire one threat GMO pathogen culture stock? Summarizing the 

principle methods from Section III, Technical Facts: 

Selection and genetic modification of organisms to create an initial pathogen tock can be done 

with relative ease. Alternative approaches to acquiring an initial stock are: 

 * Harvest from nature. By going to the origins of a pathogen, with the aid of a clinician, 

one can harvest “wild type” pathogens that are endemic to certain regions. 

 * Obtain it from a research center where work is being performed. 

 * Create it either modifying another pathogen or synthesizing it from its obtainable 

components using conventional gene-splicing techniques either in available third-party facilities 

or own dedicated facilities. This includes, at the current state-of-the art, outsourcing to DNA 

sequencing companies (also referred to as DNA foundries). 

 * By synthetic biological or abiotic techniques. 

A significant complication is the likelihood that a nation state may mask its actions and 

involvement in a given attack by equipping surrogates such as individuals or small terrorist 

groups with the means to carry out a more sophisticated attack, as Iran is thought to do with 

conventional weapons and Hezbollah. 

 3. What does it take to embed the pathogen in a medium suitable for storage and 

dissemination without killing or disabling would-be attackers and without attracting the attention 

of police and intelligence agencies? 

 4. What are the means of delivery/dissemination? 

 1. Dispersion (aerosol; FEDEX, grocery deliver system, etc.) 

 2. Infection 

 3. Poisoning 

 4. Vectors (fleas, ticks, etc) “Reservoir Vectors” that give persistence – food, 

sewage, but infrequently humans 

 5. What are the potential GMO threat outcomes (Feasible Effect & cost-Benefit)? 
27

 * Humans. Induce large numbers of victims (setting aside the psychological effect, we 

can postulate “at rates of infectivity and virulence exceeding naturally-occurring pathogens as a 

pragmatic metric); 

 * Burdening health care systems / other economic burdens 

 * Agricultural and animal industries / Disrupt Food Chain (nutrition / Economic) 

 * Area denial / Disrupt operation of critical infrastructure 

 * Equipment contamination 
28

Limited Current Analysis

The Analytical Framework captures the complexity and uncertainty of a potential GMO threat. 

From a probability and statistics viewpoint, the combinations and permutations available in the 

model yield approximately 300,000 possible pathways and outcomes. Researching each pathway 

and outcome with full enumeration is possible, but probably not useful, and a better use of the 

framework would be to demonstrate and evaluate both historical and proposed attacks from 

literature and intelligence estimates. Nonetheless, from the effort supporting the development of 

this limited framing and analysis, the following views are offered. 
29

Preliminary Findings

Using the original set of research questions posed, we found: 

1. Primary question: What is the nature and scope of the threat, if any, posed by GMOs, 

to include the potential to develop completely de novo organisms or completely artificial abiotic 

systems? 

 * The likelihood of a completely artificial or abiotic single cell entity, much less a 

deliberate pathogen, is very small. To date, despite some published claims of an artificial lifeform, biological science is, at most, still only emulating the otherwise natural fabrication of 

living entities. 

 * Modification of existing pathogens to avoid detection, be more virulent or better 

weaponized is more likely, but probably only in the hands of nation-state or above level. Overall, 

the overhead to create/use GMOs as a military weapon is only plausible at nation-state or above 

level 

 2. What are the fundamental processes and global state of the art for creating GMOs? The 

fundamental processes for creating GMOs are reflected in the Analytical Framework. The global 

state of the art for creating GMOs is more complex, but generally due to the significant global 

increases in the field of biotechnology, the primary capabilities to at least create one GMO 

culture is widely available. 

 3. Beyond the technical means to create a GMO, what might the follow-on requirements 

for “weaponization” include? 

    * Outside the laboratory, nature tends to side with the defender since ambient 

conditions tend to kill or reduce effectiveness of GMOs. Evolutionary processes suppress manmade efforts to propagate pandemic like weapons. Nonetheless, as in nature, exceptions occur. 

Sunlight (UV); heat, cold, lack of availability of a suitable host organism, all comes into play; 

therefore: 

 * The ability of most perpetrators to manufacture scale quantities (nominally 25 

gallons) is apparent. However the final steps of pathogen stabilization and delivery will elude all 

but the very competent nation state adversary. 

4. What are the capabilities and incentives for foreign states, transnational groups, small 

terrorist groups, or individuals to attempt to develop a significant GMO threat? 

 * Although possession of a capability to develop a GMO threat is plausible by a 

non-nation actor, other than using GMO to avoid detection, there is no real advantage to do so 
30

and mounting a “catastrophic” pathogen attack is more easily accomplished without GMO 

overhead and uncertainties. 

    * The classified annex to this report includes a more detailed answer to this 

question. 

Other relevant findings include: 

 * There is a trade space for some pathogens where increased virulence will result 

in “burn-out” within a confined geographical area, that is, those susceptible to the pathogen will 

succumb quickly, while those who aren’t will be immune. The propagation of the pathogen will 

then cease unless individuals break out of the confined area and further communicate the disease 

to new areas. 

    * Identifying and preventing any GMO attack will be problematic. Unlike other 

classes of weapons (e.g., nuclear devices, artillery pieces, etc.,) the science, technology, means of 

production and delivery of GMOs are demonstrably dual use. The path necessary to produce a 

beneficial GMO for commerce is often indistinguishable from that necessary to create something 

malevolent, and the path from a beneficial to a threat GMO is short and swift. The GMO threat 

generally cannot be detected by the normal intelligence collection and analysis methods. 
31

Conclusion

We conclude that, broadly stated, peaceful scientific advances, global statistics and 

demographics of GMOs suggest that the potential for corruption of biotechnology to catastrophic 

malevolent use is considerable. At a more detailed level, we find that there are tangible 

opportunities for many potential adversaries to acquire, modify and then manufactures to scale a 

potential GMO pathogen. Further development of a modified pathogen for use in a full scale 

direct catastrophic biological attack is feasible, but the full spectrum of technologies for scale-up, 

testing, packaging, weapon production and employment will most likely require the resources of 

a nation state or comparably-resourced organization. We recommend that, in concert with 

“science-based “ analysis, further efforts to expand and utilize this analytical framework be 

undertaken to better characterize the future threat from GMOs as well as other emerging 

approaches and entities such as those derived from systems or synthetic biology and 

bioregulators 
32

References 

Arne Holst-Jensen, “Testing for genetically modified organisms (GMOs): Past present and 

future perspectives”; Biotechnology Advances, 2009 

American Journal of Law and Medicine, “FDA Regulation –An Answer to the Questions of 

Human Cloning and Germline Gene Therapy”, Boston University School of Law, 2001 

Battelle, Biotechnology Summary; U.S. Biosciences employment growth 2001-2008, Battelle, 

2010 

Battelle, Biotechnology Summary; Net Income 2009; dated 2010 

Britt Erickson, (10 February 2009). FDA Approves Drug From Transgenic Goat Milk; Chemical 

and Engineering News, 10 February 2009 

Charlotte Vendrely,” Biotechnological Production of Spider-Silk Proteins Enables New 

Applications”, Macromolecular Biosciences, volume 7, Issue 4 pages 401-409. April 10, 2007 

D. B. Gibson, et al “Creation of a bacterial cell controlled by a chemically synthesized genome”. 

Science, 2010 Jul 2; 329(5987):52-6. Epub 2010 May 20 

Defense Intelligence Agency, “Unclassified Memorandum – NCI Response to ECBC”, 9 

November 2011 

David Cyranowski, “Marmoset Model Takes Centre Stage”; Nature, 459 -523-527, 27 May 2009 

David A. Relman, “Bioterrorism – Preparing to Fight the Next War,” The New England Journal 

of Medicine 354, no. 2 (2006) 

Eric Hoffman, “Dangers of Synthetic Biology in Biofuels Production”, from report 

http://www.foe .org/healthy-people/synthetic-biology, September, 2010 

Ethel Machi and Jena Baker McNeill; “New Technologies, Future Weapons: Gene Sequencing 

and Synthetic Biology”; Homeland Security 2020; The Heritage Foundation August 2010 

Frank Grotton, “Project BioShield: Authorities, Appropriations, Acquisitions, and Issues for 

Congress”, Congressional Research Service; July 7, 2010 

G. Linshiz et al., “Recursive construction of perfect DNA molecules from imperfect 

oligonucleotides”. Molecular systems biology 4: 191; doi:10.1038/msb.2008.26] 

H.O. Smith et al., “Bacteriophage from synthetic oligonucleotides”. Proc.Natl.Acad. Sci. U.S.A. 

2003, 100 (26), 15440-15445. 
33

J.D. Watson, F.H.C. Crick, Nature Archives, “A Structure for Deoxyribose Nucleic Acid”,

Nature 171, 737-738 (1953) 

James Clives, “Global Status of Commercialized Biotech/GM Crops 2008”, ISAAA Brief 39 

James J. Valdes, Ph.D., “Abiotic Networked Threat Systems, a Platform TCTI Concept”, 2008 

James J. Valdes, Ph.D., “Transformational Countermeasures Technology Initiative”, RDECOM, 

2008 

Jay D. Keasling, Production of the anti-malarial drug precursor artemisinic acid in engineered 

yeast” Nature, 13 April 2006 

Jean V. Grace, Research Challenge: “ How to Defense Against Still-Undefined Chemical, 

Biological Attacks”; National Defense, June 2010 

J.J. Esposito et al., “Genome sequence diversity and clues to the evolution of variola (smallpox) 

virus”. Science, 2006, 313, 807-812 

Jonathan B. Tucker and Raymond A. Zilinskas, “The Promise and Perils of Synthetic Biology”; 

The New Atlantis Journal of Technology and Society; 2006 

Jose‐Luis Sagripanti, Alan J., Ramsbotham, ECBC – TR‐666 – “Global Survey of Research and Capabilities

in Genetically Engineered Organisms that could be used in Biological Warfare or Bioterrorism”, 

Edgewood Chemical and Biological Center, December 2008. 

Leroy Hood, “Finding Early Signs of Mad Cow Disease”, Molecular Systems Biology, March 

2009 

M. Garfinkel, et al., http://www.thehastingscenter.org/synthetic-biology-bioethics-briefing-book/ 

Accessed 30 August 2010 

M.F. Brink, “Developing efficient strategies for the generation of transgenic cattle which 

produce biopharmaceuticals in milk”, Theriogenology, An International Journal of animal 

Reproduction, Volume 53, Issue 1 Pages 139-148, January 2000 

Militarily Critical Technologies List, Section 4, Biomedical Technology, June 2009 

Military Critical Technologies L Section 4.1 “Host Genome Material in Virus-like Agent to 

Affect Soldier Capability”, June 2009 

Mutlu Mehmet, “QCM-based biosensors for detection of genetically modified organisms 

(GMOs”, Biochemical Engineering Journal, November 2008

Ochman and Raghavan, “Excavating the Functional Landscape of Bacterial Cells”; Science 27 

November 2009 
34

Paul Cello, “Chemical synthesis of poliovirus cDNA: Generation of infectious virus in the 

absence of natural template”. Science, 2002, 9 (297), No. 5583 

R.J. Jackson, et al. “Expression of mouse interleukin-4 by recombinant ectromelia virus 

suppresses cytolytic lymphocyte responses and overcomes genetic resistance to mousepox”,

Journal of Virology. 2001, 75 (3) 

Robert Armstrong, David Franz, James Valdes, “Biological Agents: Threat, Preparedness, 

Response and Myths”, presentation to European Commission, February 2009 

 Robert Armstrong, Dr. David Franz, Dr. James Valdes, “Bill Patrick’s Relative Aerosol Potency 

Chart, Biological Agents: Threat, Preparedness, Response and Myths”. February 2009 

Rosie S. Hails, “Genetically modified plants – the debate continues”, Institute of Virology and 

Environmental Microbiology, Oxford, UK; Tree Volume 15, 1 January 2000 

United States National Academies, “Symposium on Opportunities and Challenges in the 

Emerging Field of Synthetic Biology”, Organization for Economic Co-operation and 

Development”, OECD, Royal Society 2010 

USDA Economic Research Service, Adoption of Genetically Engineered Crops in the USA,

Table http://www.ers.usda.gov/Data/BiotechCrops/ 

V. E. Volchkov et al., “Recovery of infectious Ebola virus from complementary DN: RNA editing 

of the GP gene and viral cytotoxicit”y., Science, 2001, 291, 1965-1969.

V.R. Racaniello,et al, “Cloned Poliovirus complementary DNA is infectious in mammalian 

cells”. Science, 1981, new Series (4523), 916-919 
35

Annex A. Terms and Definitions39

Fundamental Genomics – where research is generally directed towards the basic structure of 

human and microbial systems. Understanding the basic structure, organization, and function of 

human and microbial genomics potentially allows the development of genetically modified threat 

organisms resistant or immune to the natural defenses of the human body. 

Functional Genomics – Once the structure of a genome is determined (sequenced), a major task 

remaining is to determine the function of each of the genes. 

Proteomics – The complex set of proteins encoded by the cell during its lifetime is referred to as 

the proteome. On goal of genomics is to understand in detail, how the different genes encode for 

the synthesis (also known as expression) and assembly of proteins. For example, controlling the 

protein expression of antigenic proteins in a microorganism could make such a microorganism 

resistant to vaccines or undetectable by antibody-based diagnostics. Also, GMOs could be 

tailored to either express proteins or metabolize products that would be toxic to the host. 

The underlying technologies for developing and exploiting genomic information can be divided 

into broad areas that support the three sub-fields of genomics described above. 

Gene Sequencing – the basic “pick and shovel” work of determining the physical structure of 

the genome. The rate of gene sequencing and number of entities sequences has undergone 

hyperbolic expansion in the last ten years. 

Molecular biology and chemistry – including technologies for rapid screening and 

combinatorial chemistry, are essential to advancing functional genomics, and to understand the 

biological and chemical effects of specific proteins and other bio chemicals expressed during 

biological processes. 

Protein engineering and bioprocess engineering extend molecular and genetic knowledge to 

optimize large scale and affordable production of organisms and biological materials. 

Bioinformatics is a critical field comprised of state-of-the-art and entirely new information 

processing capabilities which are required to make effective use of the volume of data produced 

bio biomedical research. Bioinformatices can identify metabolic pathways from specific gene to 

end product, molecular structure, and even correlate gene sequences of threat viruses to the 

disease that they may cause. The informatics tools for systems biology, including databases such 

as Genbank and the Kyoto Encyclopedia of Genes and Genomes and analytical tools such as 

Cytoscape, now make possible the ready visualization and analysis of virtually any set of 

pathways, natural or contrived. 

Definitions exist. What they all have in common, however, is that they see synthetic biology as 

the design and construction of new biological functions and systems not found in nature.40 More 

                                                           

39 Jose‐Luis Sagripanti, Alan J. Ramsbotham, Jr. “Global Survey of Research and Capabilities in Genetically

Engineered Organisms that could be used in biological Warfare of Bioterrorism”, December 2008
36

precisely, synthetic biology seeks to mimic living systems by re-designing cells and tissues. Such 

approaches are bound by the limitations of cellular biology 41 As an example, genes, proteins, 

and functionalities are becoming increasingly fungible real-world entities that are being 

engineered as synthetic biology ‘parts,’ such as BioBricksTM “Synthetic biology may be 

especially powerful in this respect because it frees the design of biological systems for the 

process of natural evolution. The ability to sequence and then synthesize DNA (and even to 

invent new base code) adds a new layer to the power of nature: giving Synthetic Biology is a 

new area of biological research that combines science and engineering and encompasses a 

variety of different approaches, methodologies and disciplines, and many different humans the 

ability to design and redesign the biological systems of which they themselves are part.”42

Systems Biology Systems biology is the integration of the many different levels of knowledge 

(genomics, proteomics, metabolomics) about cells and organisms to gain a global understanding 

of function 

Most recently, the term synthetic biology has been adopted by an engineering cohort to define 

the process by which natural biological molecules (enzymes, DNA, proteins, etc.) are extracted 

from living systems and defined as basic building blocks to be reassembled in unnatural order 

and environments to create novel “devices or machines” that perform specific, predictable 

functions which may or may not be found in natural biological systems. This engineering 

approach differs significantly from “systems biology”, in that the individual biological constructs 

most suited to constructing a device are those units that act independently in contributing to the 

whole: the whole can be predicted from the sum of its individual parts.43

Abiotic cells include “organotypic” approaches which abstract the functionalities of living 

systems without copying their components44

Bioregulators include a variety of neurupeptides which mediate many biological functions such 

as reproduction, metabolism, growth, temperature, heart rate, behavior, memory and emotional 

state. Examples include endorphins, enkephalins, and tachykinins.45

                                                                                                                                                                                                40 Wikipedia, general description of Synthetic Biology for laymen. 41 James. J. Valdes, Ph.D. Scientific Advisor for Biotechnology (ST) “Transformational Countermeasures Technology

Initiative”, RDECOM 2008 42 U.S. National Academies Organisation for Economic C‐Operation and Development (OECD), “Opportunities and

Challenges in the Emerging Field of Synthetic Biology”, 10 July 2009 43 James Valdes, Ph.D “ANTS‐TCTI” White Paper, 44 James Valdes, Jr. Ph.D., TCTI, RDECOM proposal, 2008 45 Norbert Herzog, Ph.D, Biotechnology, biodefense and nanotechnology Advances in Academia and Industry,

Department of Pathology, University of Texas Medical Branch, TX
37

Annex B Recommendations

1. Investments. Given the findings from the overall study analysis and framework, it would be 

difficult/inappropriate to make specific investment recommendations in any particular field of 

technology or application. If there is a need for an investment determination, then in conjunction 

with a well defined set of assumptions and conditions, the framework should be utilized to make 

better informed decisions. 

2. We recommend that the basis of future GMO or related bioware studies be shifted to a more 

science-based approach. In addition to the overall complexity, uncertainty and speed of change 

with respect to biotechnology, there are significant impediments with the efficacy of traditional 

threat analysis when used in this field. 

3. In particular, standard methods of intelligence collection and analysis appear to be mostly 

based on literature and spoken intent, rather than technical competencies. Granted, there are 

fundamental difficulties to discern malevolent capabilities from the inherent dual-use nature of 

biotechnical development. Still, shifting towards a more science-based analysis, such as use of 

proxy technical means to test for key biowarfare processes, may better illuminate what the real 

threat is. 

4. The analytical framework should be expanded to include the next “horizon” of potential 

biologically based threats, such as bio-regulators or potential threats brought about through 

systems biology. An additional area on new interest would include the recent development of 

microorganisms which can attack materials, such as silicon, which may produce a new category 

of “executable” biowarfare. 

5. The results of any specific application of the frameworks towards a particular threat or context 

should be used to help update and inform the intelligence community. There is some strong 

potential that better warnings, indicators and collection methods might benefit from such 

analysis.