I-Corps: Unlocking constraints on mammalian cell programming using synthetic DNA and evolutionary selection

I-Corps项目的更广泛影响/商业潜力是开发基因工程哺乳动物细胞系，使细胞能够在缺乏特定营养物质或存在有毒废物等环境压力因素下生长。这些细胞可以降低以哺乳动物细胞为基础的制造的成本，这种制造被用于合成对现代医学至关重要的产品，如生物制剂和细胞疗法，以及可能减少全球农业碳足迹的产品的制造，如养殖肉类。例如，目前的技术无法使人造肉的成本与传统肉持平，因为人造肉95%以上的成本来自在培养基中添加重组生产的生长因子以促进细胞增殖的成本。为培养肉而培养的哺乳动物细胞可以被改造成产生自己的生长因子，以降低成本。哺乳动物细胞的基因工程复原力有可能使进行大规模哺乳动物细胞培养的成本降低到目前成本的一小部分。这个I-Corps项目是基于哺乳动物细胞编程的基因工程平台的开发。最先进的技术使DNA仅以10,000碱基对的规模写入哺乳动物基因组。这项提议的技术可能允许基因工程师将10万个碱基对写入哺乳动物细胞。这种能力可能允许编码更复杂的指令集，这些指令集可以指定复杂的细胞行为，并克服生物编程中现有的尺寸限制。此外，所提出的技术可用于解决知识约束，即从几乎无限数量的等（非）可能的潜在解决方案中选择来设计特定的行为。该平台旨在快速建立原型并并行测试数千种解决方案。这项工作是基于证明哺乳动物细胞在缺乏缬氨酸（一种典型的必需氨基酸）的情况下能够生长的技术结果。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this I-Corps project is the development of genetically engineered mammalian cell lines that enable cells to grow in the presence of environmental stressors such as the lack of specific nutrients or in the presence of toxic waste products. Such cells may lower the costs of mammalian cell-based manufacturing, which is used in the synthesis of products that are critical to modern medicine such as biologics and cellular therapies as well as in the manufacturing of products that may reduce the carbon footprint of global agriculture such as cultivated meat. For example, cost parity of cultivated meat with conventional meat is not attainable with current technologies as over 95% of the cost of cultivated meat comes from the cost of adding recombinantly-produced growth factors to the culture medium to promote cell proliferation. Mammalian cells grown for cultured meat may be engineered to produce their own growth factors to reduce cost. Genetically engineering resilience in mammalian cells has the potential to bring the cost of performing large-scale mammalian cell culture down to a fraction of present-day cost.This I-Corps project is based on the development of a genetic engineering platform for mammalian cell programming. State-of-the-art technologies enable DNA writing into mammalian genomes at only the 10,000 base-pair scale. The proposed technology may allow genetic engineers to write 100,000 base-pairs into mammalian cells. This ability may allow the encoding of more complex sets of instructions that can specify sophisticated cellular behaviors and overcome the existing size constraints on biological programming. In addition, the proposed technology may be used to solve the knowledge constraint, i.e., selecting from an almost infinite number of equally (un)likely potential solutions to engineer a particular behavior. The platform is designed to rapidly prototype and test thousands of solutions in parallel. This work is based on technical results demonstrating the ability to enable mammalian cells to grow in the absence of valine, a canonically essential amino acid.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.