The Genome Project–Write Issues, Enabling Inclusive Decision-Making on the Topics Mentioned Above (7)

Human chromosomes and nucleus as visualized by scanning electron microscopy.

Through open and ongoing dialogue, common goals can be identified. Informed consent must take local and regional val- ues into account and enable true decision- making on particularly sensitive use of cells and DNA from certain sources. Finally, the highest biosafety standards should guide project work and safety for lab workers and research participants, and ecosystems should pervade the design process. A prior- ity will be cost reduction of both genome engineering and testing tools to aid in equi- POLICY FORUM table distribution of benefits—e.g., enabling research on crop plants and infectious agents and vectors in developing nations. GENOME ENGINEERING To ensure responsible innovation and ongoing consideration of ELSI, a percentage of all research funds could be dedicated to these The Genome Project–Write issues, enabling inclusive decision-making on the topics mentioned above (7). In addi- We need technology and an ethical framework tion, there should be equitable distribution

of any early and future benefits in view of on July 29, 2016 for genome-scale engineering diverse and pressing needs in different global regions. The broad scope and novelty of the By Jef D. Boeke,* George Church,* large-scale synthesis and editing of genomes. project call for consideration of appropriate Andrew Hessel,* Nancy J. Kelley,* Adam To this end, we propose the Human Genome regulation alongside development of the sci- Arkin, Yizhi Cai, Rob Carlson, Aravinda Project–Write (HGP-write), named to honor ence and societal debates. National and in- Chakravarti, Virginia W. Cornish, Liam HGP-read but embracing synthesis of all ternational laws and regulations differ, and Holt, Farren J. Isaacs, Todd Kuiken, Marc large genomes. as in stem cell research, a major burden of Lajoie, Tracy Lessor, Jeantine Lunshof, responsibility for setting standards rests with Matthew T. Maurano, Leslie A. Mitchell, RESPONSIBLE INNOVATION the scientists and their community. Existing Jasper Rine, Susan Rosser, Neville E. Genome synthesis is a logical extension of stem cell research guidelines (8) may serve as Sanjana, Pamela A. Silver, David Valle, the genetic engineering tools that have been a useful template. Harris Wang, Jeffrey C. Way, Luhan Yang used safely within the biotech industry for http://science.sciencemag.org/ ~40 years and have provided important so- FROM OBSERVATION TO ACTION he Human Genome Project (“HGP- cietal benefits. However, recent technologi- The primary goal of HGP-write is to reduce read”), nominally completed in 2004, cal advancements—e.g., standardized gene the costs of engineering and testing large ge- aimed to sequence the human genome parts, whole-genome synthesis, and clustered nomes (0.1 billion to 100 billion base pairs) and to improve the technology, cost, regularly interspaced short palindromic re- in cell lines to 1/1000th of previous efforts and quality of DNA sequencing (1, peats (CRISPR)–Cas9 genome editing tech- within 10 years. This will include whole-ge-

2). It was biology’s first genome-scale nology (3, 4)—are revolutionizing the field nome engineering of human cell lines and Downloaded from Tproject and at the time was considered con- (5). Some applications are controversial; other organisms of agricultural and public troversial by some. Now, it is recognized as human germline editing in particular has health significance or those needed to inter- one of the great feats of exploration, one that raised intense moral debate (6). As human pret human biological functions—i.e., gene has revolutionized science and medicine. genome-scale synthesis appears increasingly regulation, genetic diseases, and evolution- Although sequencing, analyzing, and edit- feasible, a coordinated scientific effort to un- ary processes. ing DNA continue to advance at a breakneck derstand, discuss, and apply large-genome This goal is necessarily ambitious, be- pace, the capability for constructing DNA se- engineering technologies is timely. HGP- cause building a human genome at today’s quences in cells is mostly limited to a small write will require public involvement and prices would cost more than HGP-read (9) number of short segments, which restricts consideration of ethical, legal, and social im- (see fig. S1). However, an expectation of the ability to manipulate and understand plications (ELSI) from the start. Responsible HGP-write is that a sharp price drop will be biological systems. Further understand- innovation requires more than ELSI, though, catalyzed as new technology development CE U R ing of genetic blueprints could come from and involves identifying common goals im- occurs apace with advancement of the proj- O construction of large, gigabase (Gb)–sized portant to scientists and the wider public ect, as with the cost of DNA sequencing in CI E N CE S S

animal and plant genomes, including the hu- through timely and detailed consultation HGP-read. Small viral (10) and bacterial (11) / man genome, which would, in turn, drive de- among diverse stakeholders. genomes synthesized from scratch and or- Y R E D velopment of tools and methods to facilitate We will enable broad public discourse ganisms with recoded genomes (12) derived on HGP-write; having such conversations from large-scale genome editing (13) have AND S

well in advance of project implementation demonstrated the feasibility and utility of W E R The list of author affiliations is available in the supplementary O : P materials. *These authors contributed equally to this work. will guide emerging capabilities in science synthetic genomes. By focusing on building T

Email: [email protected] and contribute to societal decision-making. the 3 Gb of human DNA, HGP-write would P H O

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Published by AAAS INSIGHTS push current conceptual and technical lim- yeast genome) (14, 15), HGP-write would be makes whole- or partial-genome synthesis an its by orders of magnitude and deliver im- conducted in phases with explicit goals and efficient route to these goals. portant scientific advances. metrics. Each of the earlier large-scale proj- HGP-write will aim to address a number ects began with pilot projects focused on a PROJECT LAUNCH AND ADMINISTRATION of human health challenges. Potential ap- fraction of the genome, typically ~1%. For The goal is to launch HGP-write in 2016 plications include growing transplantable HGP-write, the pilots should provide re- with $100 million in committed support, human organs; engineering immunity to sources of immediate value for advanced from public, private, philanthropic, indus- viruses in cell lines via genome-wide recod- biomedical research and/or biotech devel- try, and academic sources from around the ing (12); engineering cancer resistance into opment. Technology development will likely world. The costs of the project lie not only new therapeutic cell lines; and accelerating also occur early in the project to propel large- in obtaining de novo synthesized DNA but high-productivity, cost-efficient vaccine and scale genome design and engineering. in the assembly, integration, and functional pharmaceutical development by using hu- A series of pilot projects making use of assays required to evaluate and understand man cells and organoids. The project could very long DNA sequences that are nonethe- the modified genomes. Total project costs are encourage broad intellectual property ac- less short of a full genome are anticipated: difficult to estimate but would likely be less cess via patent pooling. Extreme cost-re- (i) synthesizing “full” gene loci with accom- than the $3 billion cost of HGP-read. duction is feasible, as demonstrated by the panying noncoding DNA to help explain HGP-write could be implemented through $1000 genome grant program (2), as well as still-enigmatic roles of noncoding DNA one or more centers [similar to Centers of variants in regulating gene expression and Excellence in Genomic Science (CEGS) and leading to more comprehensive models for the Brain Research through Advancing Inno- Ultrasafe cell lines the role of noncoding genetic variation in vative Neurotechnologies (BRAIN) initiative Some properties of “ultrasafe” cells with a perva- common human diseases and traits; (ii) centers] that will coordinate and support for- sively reengineered genome. constructing specific chromosomes—e.g., mation and work of multi-institutional and chromosome 21—or complex cancer geno- interdisciplinary research teams working in Virus resistance types to more comprehensively model hu- a highly integrated fashion responsive to and Can be conferred by systematically recoding

man disease; (iii) producing specialized engaged with a broad public outreach. on July 29, 2016 certain codons across all genes. Subsequent chromosomes encoding one or several We celebrate 2016—the 25th anniversary deletion of tRNA genes would generate a cell line pathways—e.g., all genes needed to make of HGP-read—as a major step forward for hu- resistant to viruses. a prototrophic human cell or pathways to man knowledge and health. In this spirit, we Improved cancer resistance transform the pig genome to make it more look forward to the launch of HGP-write. Tumor suppressor genes could be made multicopy; amenable as a source for human organ REFERENCES AND NOTES genes like p53 could be recoded to eliminate CpG transplantation; (iv) a potential transfor- 1. International Human Genome Sequencing Consortium, dinucleotides that give rise to “hotspot” mutations. mation of gene therapy, with freedom to Nature 431, 931 (2004). deliver many genes and control circuits to 2. C. W. Fuller et al., Nat. Biotechnol. 27,1013 (2009). Other useful traits improve safety and efficacy, provided that 3. L. Cong et al., Science 339, 819 (2013). Delete potentially deleterious genes such delivery challenges can be met. Indeed, 4. P. Mali et al., Science 339, 823 (2013). as prion genes. 5. A. D. Haimovich, P. Muir, F. J. Isaacs, Nat. Rev. Genet. 16, 501 many substantial and useful innovations (2015). Improved genome stability may be realized in such “stepping-stone” 6. D. Baltimore et al., Science 348, 36 (2015). http://science.sciencemag.org/ projects that are short of whole-genome 7. Presidential Commission for the Study of Bioethical Comprehensively eliminate endogenous repetitive Issues, New Directions: The Ethics of Synthetic Biology and “self sh DNA” elements. reengineering but require substantial im- Emerging Technologies (Washington, DC, 2010). provement in synthesis capacity of Mb- to 8. J. Kimmelman et al., Nature 533, 311 (2016). Fail-safe security Gb-sized DNA. Both genome-wide and 9. P. A. Carr, G. M. Church, Nat. Biotechnol. 27, 1151 (2009). 10. J. Cello, A. V. Paul, E. Wimmer, Science 297, 1016 (2002). Prevent formation of germ cells, e.g., more modest changes could be tested for by removing transcriptional regulators. 11. D. G. Gibson et al., Science 329, 52 (2010). their impact on, e.g., organoid development 12. M. J. Lajoie et al., Science 342, 357 (2013).

Applications and function in vitro, facilitated by ongo- 13. F. J. Isaacs et al., Science 333, 348 (2011). Downloaded from 14. N. Annaluru et al., Science 344, 55 (2014). “Go to” cell line(s) for stem cell therapies; ing progress in stem cell differentiation and 15. J. S. Dymond et al., Nature 477, 471 (2011). robust production of biologics. “organ-on-a-chip” technologies. Novel cell 16. K. Takahashi, S. Yamanaka, Cell 126, 663 (2006). culture technologies may, in some cases, be ACKNOWLEDGMENTS sharing of CRISPR tools from over 80 labs many times more cost-effective and accu- This paper is the result of meetings held at NYU Langone through www.addgene.org. Furthermore, rate than current whole-organism testing. Medical Center 31 October 2015 and Harvard Medical School because DNA synthesis, like sequencing and Additional pilot projects being considered on 10 May 2016. F.J.I. is a cofounder of enEvolv, Inc. G.C. has computation, is foundational technology, include (v) using induced pluripotent stem financial relationships with Gen9, Editas, enEvolv, and Egenesis (companies directly related to this article; for a full list of G.C.’s HGP-write could also facilitate biological en- cells (16) to construct an “ultrasafe” human financial relationships, see arep.med.harvard.edu/gmc/tech. gineering of many organisms, accelerating cell line via comprehensive recoding of pro- html). J.D.B. is on the board of directors of Neochromosome, research and development (R&D) across a tein-coding regions and deletion of corre- Inc., and owns stock in Recombinetics, Inc., and Sample6, Inc. A.H. has investments in Autodesk, Inc. G.C. is an inventor on broad spectrum of life sciences and support- sponding genome features to increase safety patents and patent applications filed by Harvard Medical School ing basic R&D of new bio-based therapies, of such a cell line (see the box); and (vi) devel- that cover synthesis, assembly, and testing of large DNAs. This vaccines, materials, energy sources, disease oping a homozygous reference genome bear- Policy Forum is the opinion of the authors and not that of their vector control, and nutrition. ing the most common pan-human allele (or employers or institutions. The authors gratefully acknowledge the financial support of Autodesk, sponsor of the meetings. allele ancestral to a given human population) PILOT PROJECTS at each position to develop cells powered by SUPPLEMENTARY MATERIALS Similarly to other Gb-scale genomic projects, “baseline” human genomes. Comparison with www.sciencemag.org/content/353/6295/126/suppl/DC1 including HGP-read, ENCODE (which aims this baseline will aid in dissecting complex Published online 2 June 2016 to map genome functional elements), and phenotypes, such as disease susceptibility. Sc2.0 (which is synthesizing a heavily edited The pervasive nature of the required changes 10.1126/science.aaf6850

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Published by AAAS The Genome Project-Write Jef D. Boeke, George Church, Andrew Hessel, Nancy J. Kelley, Adam Arkin, Yizhi Cai, Rob Carlson, Aravinda Chakravarti, Virginia W. Cornish, Liam Holt, Farren J. Isaacs, Todd Kuiken, Marc Lajoie, Tracy Lessor, Jeantine Lunshof, Matthew T. Maurano, Leslie A. Mitchell, Jasper Rine, Susan Rosser, Neville E. Sanjana, Pamela A. Silver, David Valle, Harris Wang, Jeffrey C. Way and Luhan Yang (June 2, 2016) Science 353 (6295), 126-127. [doi: 10.1126/science.aaf6850] originally published online June 2, 2016 Editor's Summary

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