ANIMAL CLONING AND GENETIC MODIFICATION: A PROSPECTIVE STUDY
Report 1 to
Institute for Prospective Technological Studies (IPTS)
Seville
July 2005
R&D and commercialisation activities
Bruce Whitelaw1
Cecilia Oram2
Chris Warkup2
Ann Bruce3
1 Roslin Institute, Roslin, Midlothian, Scotland
2 Genesis Faraday Partnership, Roslin BioCentre, Roslin, Midlothian, Scotland
3 Innogen (ESRC Centre for Social and Economic Research on Innovation in Genomics), University of Edinburgh, High School Yards, Edinburgh, Scotland
Acknowledgements
We would like to thank Moyra Forrest from Innogen for her help with the bibliographic review, Catherine Lyall from Innogen for her help with searching literature and Eileen Mothersole for contributing her secretarial skills. In addition we would like to thank all respondents of the survey and interviewees contacted in the context of the project. Prof John Woolliams of Roslin Institute and Dr Alan Tinch are thanked for their assistance with the economic modelling related to cloning in pigs.
List of contents
Executive Summary
SECTION 1 INTRODUCTION
1.1 Introduction
SECTION 2 OVERVIEW OF RESEARCH ACTIVITIES WORLDWIDE
2.1 Introduction
2.2 Technical background
2.2.1 Methods of producing cloned animals
2.2.2 Methods of producing GM animals
2.2.3 Methods of producing GM and cloned animals
2.2.4 Summary: State-of-the-art
2.2.5 Bibliometric analysis
2.2.6 Cloning
2.2.7 GM
2.2.8 Cloning and GM
2.3 Future vision
2.3.1 ES cells
2.3.2 What species and to what applications?
2.4 Public versus private research
2.5 Main drivers for research
SECTION 3: OVERVIEW OF COMMERCIALISATION ACTIVITIES WORLDWIDE
3.1 Introduction
3.2 Methodology
3.3 Inventory of the companies active in the GM and/or cloned animal sector
3.4 Summary of products in the pipeline
3.4.1 Food production
3.4.2 Molecular pharming
3.4.3 Xenotransplantation
3.4.4 Pet sector
3.4.5 Sporting animals
3.4.6 Endangered species
3.4.7 Other possible applications
3.5 Comparison of economic structure of industry sectors
3.5.1 Food production
3.5.2 Molecular pharming
3.5.3 Xenotransplantation
3.5.4 Pet sector
3.5.5 Sporting animals
3.5.6 Endangered species
SECTION 4 TECHNO-ECONOMIC BARRIERS TO GM AND CLONING
4.1 Main technical barriers
4.2 Main barriers to commercialisation
4.2.1 Cloning
4.2.2 GM
SECTION 5 COMPARISON OF EU WITH NON-EU COMPETITORS
5.1 Current research activities in the EU
5.1.1 Summary of EU position
5.1.2 Summary of international perspective
5.2 Current commercialisation activities in the EU
5.3 Non-EU countries closest to commercialising GM and cloned animals
SECTION 6 REFERENCES
6.1 References
6.2 Bibliography
SECTION 7 GLOSSARY
SECTION 8 APPENDICES
8.1 Appendix 1
8.2 Appendix 2
8.3 Appendix 3
8.4 Appendix 4
List of figures
Figure 1: Publications relating to cloning in mammals (excluding mice), birds and fish
Figure 2: Geographical distribution of publications relating to cloning in mammals (excluding mice), birds and fish
Figure 3: Publications relating to GM in mammals (excluding mice), birds and fish
Figure 4: Geographical distribution of publications relating to GM in mammals (excluding mice), birds and fish
Figure 5: GM and cloned animals for food production in the pipeline for three different time periods
Figure 6: GM and cloned animals for pharmaceutical production in the pipeline for three different time periods
List of tables
Table 1: Chronological History of cloning in different species
Table 2: Dates of technological achievements in cloning and GM
Table 3: First reports of GM livestock produced using cloning
Table 4: Inventory of companies active in the GM and/or cloned animal sector
Table 5: Description of GM and/or cloned animals for food production
Table 6: Annual yields of recombinant protein from various sources
Table 7: Description of GM and/or cloned animals for pharmaceutical production
Table 8: Description of GM and/or cloned animals for xenotransplantation
Table 9: Description of GM and/or cloned animals for pets
Table 10: Description of GM and/or cloned animals for sporting purposes
Table 11: Key research priorities in cloning and GM
Executive Summary
The aim of this report is to summarise research and commercialisation activities in cloned and GM animals worldwide. This report considers the technical and economic challenges facing these developments as well as the relative competitive position of the EU, in so far as this can be determined.
Research
The first mammalian species cloned using nuclear transfer was sheep, in 1996. Since then several species have now been cloned including cow, goat, pig, horse, cat and most recently dog. The most research, as measured by publications, has been on cattle. The second most studied species is the pig, mostly for xenotransplantation applications, where cloning is combined with genetic modification. The first genetically modified (GM) mammal was produced in 1985 and GM pigs, sheep, cattle, goats, rabbits, chickens and fish have all been reported. The main research use of GM techniques, as measured by publications has also been for xenotransplantation.
A GM animal is one that has had ‘new’ DNA added to its germline. The introduction of new genetic material can range from the addition of a given sequence to the replacement of a given target sequence. Delivery of the new gene (or ‘transgene’) can be achieved by direct injection or viral infection. With each approach the target can be the oocyte, sperm or fertilised egg.
Cloned animals can be produced by nuclear transfer and efficiency of cloning has been improved by several variations on this technique. Animals that are both cloned and GM can be produced by the insertion of genetic material into a cell either by direct injection or using viruses, followed by nuclear transfer from that cell to produce a clone. The first report of a genetically modified and cloned animal (sheep) was in 1997. Much of the research in cloned and GM animals has been accomplished by commercial establishments.
GM and cloning research can significantly advance our understanding of biology as well as inform animal applications more directly. Research with animal embryonic stem cells and cloning can be expected for example to inform surgical use of stem cells in human medicine. Thus research on cloning and GM in animals can be viewed as underpinning important developments in human health.
Commercialisation
The main potential commercial applications of cloned and GM animals include: food production, production of pharmaceuticals (‘pharming’), xenotransplantation, pets, sporting animals and endangered species. Of the 35 companies worldwide identified as working with GM or/and cloned animals at the time of a survey undertaken for this report; 40% were working in ‘pharming’, 15% in food production, 12.5% in pets, 10% in xenotransplantation, 10% in endangered species and 7.5% in other activities.
Cloned or GM animals already on sale include cloned pet cats, GM ornamental fish, cloned horses and at least one rodeo bull. Individuals from some endangered species have been cloned e.g. gaur, mouflon, banteng and African wildcat and cloning technology has been applied to restoring endangered breeds of cattle.
A few new products are estimated to be at or near market and are likely to be commercially available within the next 5 years, on the basis of evaluation by the commercial companies themselves. Two pharmaceutical products from the milk of GM animals have completed (Phase III and Phase II) clinical trials respectively and may be on the market in the EU in the next few years.
Cloned livestock (especially pigs and cattle) are widely expected to be used within the food chain somewhere in the world before 2010. It is likely that, within this timescale, it would not be economic for cloned animals to be used directly for food or milk production, but that clones would be used as parents of slaughter pigs, beef cattle and possibly also milk-producing dairy cows.
Faster-growing GM salmon are awaiting regulatory approval, principally for direct sale to fish farming markets in N. America, Asia and S. America.
In the longer-term (beyond 2010) potential developments include the following:
Food production: GM fish other than salmon. For terrestrial livestock, a number of potential applications include improving animal product composition, improved disease resistance, reduced environmental impact and sterility.
Molecular pharming: A range of proteins produced from GM or GM and cloned cattle, goats and chickens. Recombinant human mono- and polyclonal antibodies may be produced by GM livestock beyond 2015.
Xenotransplantation: Proponents suggest that GM pig organ xenotransplantation to humans is at least 10 years away
Technical barriers
Cloning
The main challenge for cloning animals is to improve efficiency and overcome problems of Large Offspring Syndrome. Progress has been made with respect to both these, with some commercial sources questioning the need for further improvement based on their internal data for livestock species.
Cloning dogs proved more difficult than cloning livestock, but has now been achieved. Cloning endangered species is difficult due to their unknown reproductive physiology and difficulties in providing a reliable oocyte maturation environment. The lack of availability of suitable recipients also constitutes a barrier.
GM
Major technical challenges with respect to GM include improving the accuracy of incorporating transgenes (to avoid impacts on other parts of the genome or multiple copies of genes) and improving efficiency. For GM fish, the lack of suitable regulatory elements to control transgene expression is a challenge.
A major technical barrier in the food sector application of GM technology is the time required to incorporate GM animals into existing breeding schemes. One of the technological barriers to commercialisation of GM fish is sterilization efficiency, currently below 100%, which is important if the fish are to be farmed in sea cages.
Technical barriers in the pharming sector include the long gestation period in cattle and the limited number of antibodies which can be produced. Production of pharmaceuticals in chicken eggs has been limited as the techniques for genetically modifying chickens have proved to be more difficult.
Xenotransplantation remains a significant technical challenge, not least because multiple transgenes are needed in order to overcome immune rejection mechanisms.
GM and cloned
Specific technical challenges for GM and cloned animals include the effects of ageing on donor cells before nuclear transfer can be carried out and eliminating the need to include a selection marker gene during the genetic modification step.
Main barriers for commercialisation
Cloning
The main barriers to commercialising cloning of food animals are consumer/food retailer acceptance and, particularly in the USA, the regulatory uncertainty which currently exists. The regulatory uncertainty in the USA may be resolved within the next year or so. The economic barrier to entry into animal cloning is likely to be low. Although it is not yet clear what the costs of obtaining regulatory approval will be, these costs are likely to be substantially lower than for GM livestock.
GM
The main barriers are perceived to be regulatory approval and consumer/food retailer acceptance.
Regulatory approval of pharmaceutical products from GM animals is seen as the principal hurdle followed by the time taken to get the product to market. Some of these barriers are common to all pharmaceutical products, but there may be additional uncertainties to be resolved for products from GM animals.
A barrier to attracting venture capital to start-up businesses working in this area of science is the controversial nature of the work and the complicated IP situation for cloning technology.
European competitiveness
Research
Cloning research is undertaken primarily in USA, Far-East and EU countries. Although a late entrant into the cloning research field, the Far-East may be rapidly becoming the major research player. The majority of GM research has been carried out in the USA and Europe, with Europe initially displaying a clear lead over the rest of the world. The majority of GM fish effort is in the Far East and USA. As with cloning, the Far-East is a late entrant into this area of research but is fast catching up.. If Europe aims to be a major player in cloning or GM research then it will have to focus on specific aspects of this subject and to co-ordinate its currently fragmented research effort (as is being done in the USA for example through initiatives such as the National Swine Resource Centre).
Commercialisation
We have identified 35 companies worldwide involved in producing GM animals, cloned animals or both GM and cloned animals. Of these, 63% are in N. America, 14% in Europe and 11% in Asia.
Within the EU, France is the country which seems to be most actively commercialising GM and cloned animals. French companies are applying GM and cloning to pharming, sporting and endangered animal sectors. The Netherlands and UK each have one company active in the pharming sector.
In terms of number of companies and scale of activity the USA has the most. Canada is second but with significantly less activity than the USA. We should however note that we experienced difficulty in obtaining information from companies in the Far East.
Non-EU countries are applying GM and cloning to a broad spectrum of possible application sectors. The most significant difference between EU and non-EU countries is the focus of non-EU countries on the application of GM and cloning to food production. In contrast the companies in EU countries are applying GM and cloning principally to pharming but also to sporting or endangered animals.
SECTION 1 INTRODUCTION
1.1 Introduction
Since the development of cloned animals will facilitate the production of GM animals and many of the applications of cloning (such as xenotransplantation) will depend also on the use of genetic modification, these two technologies are investigated together. The purpose of this study was to investigate the current status of developments in cloned and GM animals and more specifically to provide a comprehensive picture of R&D and commercial activities involving animal cloning and/or GM and their products world-wide; and a projected pipeline of products for the next 5-10 years. It should be stressed that, in relation to commercialisation activities, this study is very much a snapshot in time. Many of the companies involved in GM and cloning developments in animals are small companies and they operate in an area of significant commercial risk.
Regulatory, trade and socio-economic aspects are further considered in reports 2 and 3.
Cloned and GM animals can be used for a wide range of applications including food production, molecular pharming, xenotransplantation, the pet sector, sporting animals and endangered species. Development of animals to model human diseases and to investigate gene function in model animals have however been specifically excluded from this report. Within this report the term ‘cloning’ should be understood to mean somatic cell nuclear transfer (SCNT) unless otherwise stated and the term ‘animal’ should be understood to refer to non-human animals.
This report is in five sections:
- Section 1 is an introduction
- Section 2 maps research and development activities worldwide involving cloned and GM animals
- Section 3 maps out commercialisation activities.
- Section 4 considers the technical and economic barriers to developments with respect to cloned and GM animals and finally;
- Section 5 considers the position of the EU in comparison to non-EU competitors.
This study was conducted March-July 2005. The scientific aspects were investigated by staff at Roslin Institute, the commercialisation activities by staff at Genesis Faraday Partnership and co-ordination was provided by the Innogen Centre. The methodology consisted of literature and web surveys supplemented by interviews, bibliometric analysis based on Articlefirst and Web of Science, and questionnaires, as outlined in each of the relevant sections. Additionally, useful interchange of information took place with the Specific Support Action “Farm animal cloning and the public” and in a 2-day workshop held in Seville in June 2005, co-organised by the IPTS and the above mentioned SSA.
SECTION 2 OVERVIEW OF RESEARCH ACTIVITIES WORLDWIDE
2.1 Introduction
This section aims to convey an overview of on-going research activities and technological developmentsin animal cloning and/or genetic modification worldwide, including national and EU funded programs. Specific emphasis is placed on short and medium term visions (5 and 10 year span).
GM and Cloning are intertwined in both public perception and application, although they represent different technical methods in animal biology. This will be described using examples and species differences. Applications in large animals are mainly restricted to commercial projects due to the cost and effort involved. This aspect will be developed through discussion of the drivers and incentives for research in this area and contrasted to the main technical barriers for research. In addition, the position the EU holds in the global arena will be presented.
This report is based on original data and quantitative information on GM and/or cloned animals, their offspring and derived products worldwide. It also builds on publicly available information in the literature, relevant databases (as outlined in Appendix 1) and expert interviews. The expert interviews were performed face to face when possible, with a few by phone, and by email. In total 34 experts in the cloning and GM field were contacted. The geographical distribution of the experts was 23 from the EC, eight from the USA and three fromAustralasia (including one from China).
To emphasise, this section deals with the research aspects of cloning and GM. By definition there is a strong focus on technical and proof-of-concept aspects of cloning and GM.
2.2 Technical background
2.2.1 Methods of producing cloned animals
Cloning in this context relates to the production of animals through transfer of the genetic material from one donor cell to a recipient unfertilised oocyte that has had its nuclear DNA removed (enucleation). This process is also known as somatic cell nuclear transfer (SCNT). Through the use of several individual cells from a given unique source and an equivalent number of recipient oocytes, several cloned animals can be produced.
i. Overview
Although in use and robust, the standard method for producing transgenic animals apart from mice is inefficient, not exact and costly. These limitations drove researchers to seek other methods. Half-a-century ago, the background to animal cloning was demonstrated by transplanting nuclei from the blastula of the frog to an enucleated oocyte, obtaining a number of normal embryos in the process. Subsequently, nuclei from various types of cell were transplanted to an oocyte that has been subjected to ultraviolet radiation to destroy the peripheral chromosomes. Again embryos were produced. These results establish the important principle that while animal cells become committed to their fate as development proceeds, the nuclei of most cells still retain all the genetic information required for the entire developmental programme and can be reprogrammed by the cytoplasm of the oocyte to recapitulate (restart) development (reviewed by Campbell et al. 1996).