FAST-STEM: Synthetic Biology Toolkit for Next Generation Regenerative Medicine

Abstract

Very recent clinical advancements in human stem cell (hPSC) based tissue replacement have opened a new frontier of possibility: therapeutically augmented human cell engraftment. Therapeutic augmentation requires the design of synthetic gene circuits that are compatible with hPSCs. Such design is highly challenging due to: (a) poor translation of circuits developed in immortalized cell lines; (b) absence of effective tools for synthetic human stem cell development. To address this, we developed FAST-STEM (Facile Accelerated Stem-cell Transgene integration with SynBio Tunable Engineering Modes), a platform that enables design–build–test–learn style hPSC engineering. FAST-STEM facilitates fast iterative synthetic cell evolution, zygosity manipulation, and highly controlled library screening. Differentiation ready synthetic stem cells can be generated an-order-of-magnitude faster by FAST-STEM than by current site-specific methods. We demonstrate FAST-STEM potential to revolutionize human synthetic biology, by creation of synthetic human liver, glial-cells, muscle, cardiomyocytes, pancreatic beta-cells, cortical neurons, and dopaminergic neurons.