SUPPORTING INFORMATION Bioart Ali K. Yetisen1,*, Joe Davis,2,3 Ahmet F. Coskun,4 George M. Church,3 Seok Hyun Yun1,5* 1 Harvard Medical School and Wellman Center for Photomedicine, Massachusetts General Hospital, 65 Landsdowne Street, Cambridge, Massachusetts 02139, USA 2 Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 3 The Department of Genetics, Harvard Medical School, Harvard University, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA. 4 Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 East California Blvd, Pasadena, California 91125, USA 5 Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA * corresponding authors: ayetisen@mgh.harvard.edu, syun@mgh.harvard.edu Contents 1. Academic Bioart Programs 2. Davis’ Projects 3. Emerging Bioart Concepts 4. Opportunistic Pathogens 5. Biopaintings 6. Emerging Technologies in Genetics 1. Academic Bioart Programs Programs include courses at San Francisco State University (1999), SymbioticA (2000) at University of Western Australia (Perth), Incubator (2009) at University of Windsor (Ontario), The Bio Art Lab (2011) at School of the Visual Arts in Chelsea, NYC., and joint courses (2014) offered by departmental faculty in both art and biology at the University of Kentucky (Lexington). Courses focusing on bioart are now taught at University of Washington (Seattle), Carnegie Mellon University (Pittsburg), State University of New York (Buffalo), Ohio State University (Columbus, OH), Rensselaer Polytechnic Institute (Troy, NY), Leiden University (The Netherlands), and the School of the Art Institute of Chicago. 2. Davis’ Projects In 2003, Davis collaborated with German Engineers Thomas Kaiser and Jens Tuchsherer to create DNAgraphy, which used DNA instead of conventional silver halides to form extremely high-resolution photographic emulsions on glass slides. Davis and Kaiser displayed their DNAgraphic Slides at the L’Art Biotech, an early bioart exhibition, held at Le Lieu Unique, in Nantes, France. Davis has also created several laboratory instruments for art and science including his Audiomicroscope (ca. 1998) and Fishing Microscope (2001). The former was exhibited at Ars Electronica in 2000 while Fishing Microscope was the feature subject of a prime-time U.S. national television broadcast (ABC Nightline) in July, 2001. Continuing interest in overlaps of art, biology and SETI research led Davis to transmit a second powerful radar signal to nearby sun-like stars in 2009 on the 35th anniversary of Frank Drake’s famous 1974 SETI transmission from Arecibo Radar in Puerto Rico. Davis’ project, Rubisco Stars, was a signal transmitted from Arecibo that was encoded with the gene for ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the most abundant protein on Earth [1]. Davis’ ongoing collaborations in art and science include work with researchers at Harvard and the National Agrosciences Institute in Japan to create stable strains of genetically altered silkworms. Davis and his collaborators have produced silkworms having fibroin (silk protein) gene fusions with a gene for silicatein, protein from marine organisms that normally polymerizes silica (glass) from seawater but that will also chelate metals. Davis and his collaborators have recently produced silks integrating precious metals (Au and Pt) and metals modeling reactor meltdown byproducts with a view to applications in environmental remediation. In another project, Davis organized an international consortium to sequence and assemble the genome of Malus sieversii, progenitor of all domestic apple species (Malus x domestica). It is perhaps the first bioart project to extensively involve sequencing technologies and bioinformatics that comprise the field of genomics and the first bioart to deal with an entire genome. Legends about a tree of temptation guarded by a serpent long predate Judeo-Christianity and while traditional and historical contexts have contributed to a profoundly poetic and artistic project, apples are also one of the most widely cultivated fruit crops. Despite agricultural value of domesticated apples, populations of the wild Malus sieversii progenitor species still exist today, closely related to those of biblical time periods. They display diverse phenotypic characters that represent a critical genetic resource for disease resistance, fruit quality, and tree physiology of today’s cultivated apple. Traits lost in domestic apples may thus be recovered. Davis’ project to resolve the genome of Malus sieversii closely interweaves objectives of art and scientific research and may represent an unprecedented approach to the practice of both. Davis and his colleagues at Harvard have also created an extremely compressed version of Wikipedia that they plan to clone in the form of DNA into Malus sieversii to create a “tree of knowledge” they are calling “Malus ecclesia” [2]. 3. Emerging Bioart Concepts Many examples of bioart entail artistic manipulations of living systems [3]. However, advances in biotechnology have provided subject matter for many works of art executed in non-biological media. These have appeared in various architectural, digital, and computational forms. For example, ‘Synthetic Aesthetics’ a joint program of University of Edinburgh and Stanford University was devoted to joining synthetic biologists, designers, artists and social scientists to explore collaborations between synthetic biology, art and design [4]. This initiative focused on biomimetic and synthetic biology solutions to practical problems in optimizing design of nextgeneration structures including household architectures. Synthetic Aesthetics teams used xylem cells for instance, to determine design principles for stronger buildings. Interfaces between biology and animation, where traditions of scientific illustration have been overtaken by technology for digital modeling and 3D printing, may also have impacts on bioart. Artists can now simulate bioart in the same way that scientists can run virtual experiments in silico to precede or replace experiments in real life. Arthur Olson’s Molecular Graphics Laboratory at Scripps Research Institute is one example, where computational analysis and modeling tools are used to elucidate biomolecular interactions for research, industry and education (Fig. S1A,B). Other examples can be found in computer graphics and animations designed by Harvard’s Robert Lue for blending online and classroom education. Figure S1. Emerging bioart concepts. (A) Scientific visualization of a bushy stunt tomato virus. Scale bar = 5 nm. Courtesy of Arthur Olson and reproduced with permission. (B) 3D printed molecular structure of a virus. Scale bar = 1 cm. Courtesy of Arthur Olson and reproduced with permission. (C) DNA origami based nanoscale face structure. Scale bar = 20 nm. Reprinted with permission from ref [5]. Copyright 2006 Macmillan Publishers Ltd. DNA nanotechnology may also foster corresponding developments in bioart. DNA molecules have been utilized directly as structural elements to form constructs for both scientific and artistic purposes. Methods pioneered by Nadrian Seeman at New York University utilize DNA molecules to form nanoscale structures and programmable materials with mechanical properties. Many such structures have been created to date, including two- and three-dimensional structures, and periodic, aperiodic, and discrete structures [6, 7]. Similarly, Paul Rothemund at the Caltech has employed ‘DNA origami,’ inspired by Japanese paper folding art, to create nanoscale shapes and devices [5]. Examples of DNA origami bioart included a range of 2/3D shapes including smileys, stars, and other geometric structures (Fig. S1C). 4. Opportunistic Pathogens Bacillus atrophaeus original and still most prominent use is as a surrogate organism for B. anthracis, the anthrax pathogen. B. atrophaeus is an easily recognized simulant since it has a black pigmentation that is detectable on culture plates. A 2004 study prepared for the U.S. National Academies described B. atrophaeus pathogenic activity: “Bacillus globigii has been called B. subtilis var niger, B. licheniformis, and most recently, B. atrophaeus is now considered a pathogen for humans [8]. Most infections are associated with the experience of invasive trauma (e.g., catheters, surgery) and/or a debilitated health state, thus it is often encountered as a nosocomial pathogen. Bacillus globigii is also a well-known cause of food poisoning, resulting in diarrhea and vomiting. Infections are rarely known to be fatal, although fatal food poisoning has been reported. Ocular infections, bacteremia, sepsis/septicemia, ventriculitis, peritonitis are reported types of infection and they are usually treated with antibiotics” [8]. S. marcescens, another bacterial strain found in Kurtz’ home, is also a classic biological warfare simulant. In a series of experiments carried out in 1950 and 1951, aerosolized S. marcescens was systematically released from U.S. Navy vessels in San Francisco Bay and monitored over inland areas to test effectiveness of dispersal methods. Kurtz claims to have been concerned with using bacteria that would not harm himself or anyone in his project audience, but the 1950-51 releases of aerosolized S. marcescens were later implicated in the death of one man and the hospitalization of 10 men and women at a Stanford University hospital. S. marcescens is a human pathogen responsible for a significant percentage of nosocomial (e.g., hospital-acquired) infections. People with compromised immune systems, such as individuals who are HIV positive, or those undergoing chemotherapy, or receiving immuno-suppressants for transplant surgery are particularly susceptible. Serratia marcescens may result in sepsis, Serratia infection, urinary tract infection, respiratory tract infections, meningitis, cerebral abscess, bacterial keratitis, and bacterial parotitis [9, 10]. Senate hearings were held in 1977, and the U.S. military was criticized for the continued use of S. marcescens following awareness of the Stanford outbreak. S. marcescens has been recognized as an opportunistic pathogen in humans since the 1960s [11]. When Kurtz became aware that S. marcescens has been associated with pneumonia and urinary tract infections, he wrote to Professor Ferrell at Carnegie Mellon asking for "any other ideas on another bacteria (sic) that can travel by air and be easily identified on a Petri (sic) dish, and most importantly, is unequivocally classified as nonpathogenic?" Kurtz decided to release both S. marcescens and B. atrophaeus. In 2005, Kurtz and Critical Art Ensemble (CAE) carried out a curious homage to a 1949 U.S. biowarfare group experiment, which released S. marcescens into air ducts at the Pentagon successfully contaminating the building and convincing frightened officials to devote more money to biowarfare defense. CAE used S. marcescens to intentionally contaminate NGBK gallery in Berlin Kreuzberg and demonstrated that contamination to gallery visitors with culture plates set out in the gallery space [12]. In 2005, Kurtz and CAE re-enacted strategies employed in a 1952-53 British military exercise undertaken to determine whether or not ship’s crews could be contaminated with plague bacillus dispensed as an aerosol spray. To carry out this re-enactment, CAE dispensed a broth containing Bacillus atrophaeus in an aerosol spray from a boat off Isle of Lewis in Scotland toward a floating platform holding 30 guinea pigs and an animal-protection supervisor. A film documenting the CAE Isle of Lewis tests was shown at New York’s Whitney Biennial in 2006 [12]. Kurtz indictment was sensationalized in the press evoking significant support for Kurtz among artists, curators and members of the scientific community. A “Critical Art Ensemble Legal Defense Fund” campaign raised over $350 K for the defense of Kurtz and Farrell [13]. The indictment for mail and wire fraud was ruled "insufficient on its face" on April 21, 2008 by the presiding Judge Richard Arcara. Even if the actions alleged in the indictment (which the judge must accept as fact) were true, they would not constitute a crime. Ferrell had already been convicted for ordering S. marcescens and B. atrophaeus under false pretenses for transfer to Kurtz, while Kurtz never ordered anything from American Type Culture Collection (ATCC). 5. Biopaintings Contributors include geneticist Hunter Cole (formerly Hunter O’Reilly), a lecturer at Loyola University Chicago's Department of Biology, who in 2005 exhibited Viewing DNA Under the Moonlight and Living Drawings Created with Bioluminescent Bacteria at Loyola University Museum of Art in Chicago [14]. Furthermore, in 2005, Jeff Tabor, a professor of bioengineering at Rice University created the Bactograph project that employed light-sensitive bacteria to create images on Petri dishes (Fig. S2A) [15]. Additionally, biophysicist Nathan Shaner created Beach Drawn 80’s Style with Fluorescent Bacteria (2006) in Roger Tsien’s laboratory at the University of California San Diego using an eight-color palette of bacterial colonies expressing fluorescent proteins (Fig. S2B). Figure S2. Biopaintings. (A) Light-sensitive bacteria that exhibited patterns when exposed to light through a mask by Jeff Tabor and Matt Good. Scale bar = 1 cm. Reprinted with permission from ref [15]. Copyright 2011 Elsevier B. V. Publishers Ltd. (B) A scene drawn with an eight color palette of bacterial colonies expressing fluorescent proteins. Scale bar = 1 cm. Courtesy of Nathan Shaner and reproduced with permission. Other artists with formal scientific or technical credentials have contributed biologically inspired work to artistic venues. Australian artist and mechanical engineer Natalie Jeremijenko's One Tree project, first displayed at the 1999 `Ecotopias' exhibition at Yerba Buena Center for the Arts in San Francisco, and later at the ‘Paradise Now’ exhibition in New York, involved use of plant tissue-culture techniques. Jeremijenko created 1000 genetically identical clones of a single `paradox tree' (an ornamental variant of the walnut tree), attempting to demonstrate that subtle environmental differences can produce large variations in the phenotype of genetically identical trees [16]. University of Montreal evolutionary biologist and artist François-Joseph Lapointe has applied his evolutionary algorithms to dance. His work, Choreogenetics (2005), was based on a fragment of the D-loop region of mitochondrial DNA (mtDNA). Lapointe first performed the piece as a solo, then as a duo (gallery performances at Place des Arts in Montreal), and finally as a group piece for 30 dancers (University Space performance at Institut National des Sciences Appliquées, Lyon, France) [17]. 6. Emerging Technologies in Genetics Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) technology now stands ready to transform the field of biology [18-20]. CRISPR is a new genome editing tool that may revolutionized the field of genetic engineering with unprecedented capabilities for precision, efficiency, and flexibility. The appearance of new technologies often comes with new sets of risks and rewards and CRISPR is no exception. CRISPR-mediated edits can modify cell germlines, allowing specific changes to be inherited. CRISPR can be used to build ‘gene drives’ causing an editing event to re-occur in each organism that inherits it and could theoretically be used to spread any trait throughout entire populations. CRISPR could theoretically alter every ecosystem on Earth [21]. As a safeguard, biologists using the same CRISPR techniques have shown that population-level genome changes could be reversed or blocked permitting a variety of gene drive intended to control rather than eliminate entire populations. In 2015, researchers successfully tested CRISPR-based yeast and Drosophila gene drives in the laboratory [22]. Bioartists may be expected to investigate many other new techniques as well, including induced pluripotent stem (iPS) cell technology [23-25]. It is possible to induce iPS cells to form whole organs in culture, a capability that may go beyond the ambitions of Mary Wollstonecraft Shelley’s ‘Victor Frankenstein’. Bioartists can of course be expected not only to contemplate these possibilities, but to create ‘living art’ or, ‘semi-living art’ based on such systems. Artists are unlikely to ignore ‘artificial life’ created with unnatural nucleotides and unnatural amino acids; new nanotechnology to visualize and interact with proteins and other biomolecules on smaller scales; advances in synthetic and systems biology, astrobiology, and neurosciences. References 1. Davis, J. and Hofmans, D. (2010) RuBisCo Stars and the Riddle of Life. LPI Contributions 1538, 5370 2. House, P. (2014) Object of Interest: Twice-forbidden Fruit, The New Yorker 3. Kac, E. (2007) Signs of life: bio art and beyond. MIT Press, Cambridge, MA 4. Ginsberg, A.D., Calvert, J., Schyfter, P., Elfick, A., and Endy, D. 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