Why Democratize Biotechnology In the coming years, rapid development and worldwide deployment of advanced biological technologies in the areas of medicine, agriculture, industrial manufacturing and ecosystem services <a href="https://newconsensus.com/projects/biotechnology-and-the-green-new-deal" target="_blank" rel="noreferrer noopener">will be needed</a>  in order to help transition human civilization off of fossil energy and materials and avert a mass extinction. Right now, we don’t have anywhere near enough bioengineers or biotechnological productive capacity for this task, in large part due to the high financial, technical and legal (intellectual property / IP) barriers to learning and practicing bioengineering. This shortage of worldwide biotechnological productive capacity has been highlighted during the COVID-19 pandemic, with large swathes of the world's population, particularly in the less affluent Global South, still unable to access diagnostic tests and vaccines after a year and a half of the novel coronavirus spreading, evolving, and ravaging humanity around the world.  As with COVID, so with climate change: a global response at the necessary scale and speed would require an expansion and worldwide distribution of the means of producing the technological and industrial measures, including biological technologies, to combat and mitigate the crisis. Who We Are We are <a href="https://hackernoon.com/how-open-source-synthetic-biology-can-make-medical-solutions-more-accessible-ns2i32lx" target="_blank" rel="noreferrer noopener">Friendzymes</a> , an international collaboration of community and academic biology labs, professional and amateur bioengineers with members from the USA, India, Ghana, the UK, Singapore, the Philippines, Brazil, Canada, Perú and the UAE. Our collaborating labs currently include BioBlaze in Chicago, led by PI Dr. Sarah Ware; Hive Bio Lab in Ghana, led by PI Harry Akligoh; Open Science Network in Vancouver, led by PI Dr. Scott Pownall; and the Phillipine Genome Center at Mindanao, led by PI Dr. Lyre Espada Murao. What We’re Doing Our goal is to democratize and distribute the means of biotechnological production. Specifically, we are working to make it as cheap and easy as possible to manufacture and purify proteins and enzymes, the atomic machines that are the workhorses of modern biotechnology. Our initial target enzymes include Pfu-Sso7d thermostable DNA polymerase, the ligases and restriction enzymes necessary to perform modern genetic assembly reactions, and insulin (in partnership with the Open Insulin Project). How We Will Accomplish This We are leveraging low-cost and open-source wetware, software, hardware and law-ware at the cutting edge of synthetic biology. With support from the <a href="https://stanford.freegenes.org/" target="_blank" rel="noreferrer noopener">Free Genes Project</a> and the International Genetically Engineered Machines (<a href="https://igem.org/Main_Page" target="_blank" rel="noreferrer noopener">iGEM</a> ) competition, we are developing libraries of genetic parts that can be used to engineer well-established industrial microorganisms, including Bacillus subtilis and Pichia pastoris, to produce and secrete our target enzymes. In partnership with the <a href="https://github.com/TimothyStiles/poly" target="_blank" rel="noreferrer noopener">Poly</a> project, we are developing open-source software tools and tutorials that make it easy to design genetic parts and constructs that are synthesizable, genetically stable and will produce large amounts of the desired enzymes. We will integrate low-cost automated liquid handling robots, nanopore DNA sequencers, and open source designs for plate readers and bioreactors into frugal, high-capacity biofoundries that enable construction and testing of large numbers of genetic constructs, and scale-up of high-producing strains to the 5 liter scale. We are making all our designs open source: software is distributed under the MIT license, and all our wetware will be off-patent and distributed under the Open Material Transfer Agreement (<a href="https://biobricks.org/open-material-transfer-agreement/" target="_blank" rel="noreferrer noopener">OpenMTA</a> ). Integrated together, these pieces of hardware, software, wetware and law-ware could constitute a new and largely open-source biotechnological infrastructure stack (a ‘biostack’) that greatly lowers the financial and technical barriers to bioengineering. Once we express and secrete proteins, we will establish protocols for the purification of enzymes using the most scalable and inexpensive approaches possible (e.g. dialysis, reversible separation with elastin-like polypeptide tags, kosmotropic salt precipitation, and frugal chromatography with silica-binding peptide tags). Friendzymes and iGEM To organize our project and raise its visibility in the bioengineering community, we are participating in this year's iGEM competition (which is how we came to make the video you saw above). In the coming months, our goal is to design and assemble a frugal enzyme production and purification process for Pfu-Sso7d from Bacillus subtilis as our first target.  We will present our progress in November 2021 at the iGEM Giant Jamboree, to approximately 6000 aspiring young scientists; engineers, and professionals in the field from all across the world, whom we will recruit to help us expand  toward our goals. Who Already Supports Us We have received generous donations from Lee Redden, Scot Bryson, Andrew Hessel, and the <a href="https://openbioeconomy.org/" target="_blank" rel="noreferrer noopener">Open Bioeconomy Lab</a> , which have enabled us to register our team in the 2021 International Genetically Engineered Machines (iGEM) competition. We have further been awarded an impact grant from iGEM and a microgrant from the Just One Giant Lab (<a href="https://jogl.io/" target="_blank" rel="noreferrer noopener">JOGL</a> ) community, enabling us to attend the iGEM Giant Jamboree and pay for some of the enzymes and labware that we need. We are the first recipients of a fiscal sponsorship program by the Gathering for Open Science Hardware (<a href="https://openhardware.science/" target="_blank" rel="noreferrer noopener">GOSH</a> ), through which our project has been granted a bank account and 501(c)3 nonprofit status in the USA, making donations to this GoFundMe tax-deductible for US citizens. Finally, an exceedingly generous sponsorship from <a href="https://opentrons.com/" target="_blank" rel="noreferrer noopener">Opentrons</a>  means that all four of our collaborating labs will have the fully-stocked OT-2 liquid handling robots that are one of the core components of our frugal biofoundry design. What We Need Thanks to iGEM and the Free Genes Project, most of the wetware libraries of synthetic DNA that we design will be made and shipped to us for free; and thanks to Opentrons, so will the liquid handling robots for our biofoundries. However, we still need to purchase equipment, plasticware and glassware, enzymes, DNA primers and other reagents in order to build both our biofoundries and genetic constructs and strains that will enable high production of our target enzymes.  Specifically, each lab will need the following:   1.  <a href="https://store.nanoporetech.com/us/minion.html" target="_blank" rel="noreferrer noopener">MinION</a>  nanopore DNA sequencer with a supply of Flongle flow cells and sample preparation kits (~$3000)   2.  Components for building an <a href="https://github.com/brianchowlab/OSP" target="_blank" rel="noreferrer noopener">open source plate reader</a>  (~$4000)   3.  Components for building an open source bioreactor (~$200)   4.  Plasticware and glassware (~$1000)   5.  Enzymes and reagents for PCR and Golden Gate assembly (~$1000)   6.  Buffers, growth media, antibiotics and other reagents for microbial cell culture (~$500)   7.  DNA synthesis (primers, some synthetic genes) (~$1000) The total funding we need to build one frugal biofoundry (minus the cost of the OT-2 liquid handling robots donated by Opentrons) is roughly $10,700; while the total funding we need to build frugal biofoundries at all four of our collaborating labs is roughly $42,800. We have set the initial fundraising goal for this GoFundMe at the total required for a single biofoundry, but if we raise that amount we will increase the goal and try to build biofoundries in all our labs. Please consider donating and helping us to democratize the means of biotechnological production, accelerate the bioengineering revolution and help to build a good future together, all around the world!

Frank Lee

Anonymous

Sarah Ware

Karl Schmieder

Arye Lipman

Xavier Coadic

Kahlil Corazo

Melanie Stegman

Hakan O.

Rick Byers

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