Collaborative Research: ProteoCell: The Fat-Free Cell

Project: Research project

Description

Overview. Compartmentalization is one of the hallmarks of life: it is a fundamental process in living cells, defining both the boundary between the cell and the environment, as well as the boundary between the cytoplasm and its subcellular organelles that drive cell function and development. In extant cells, compartmentalization is generally accomplished by membranes formed by lipid bilayers that control access to/from the cell, that sequester and segregate biochemical reactions, and that host structures governing communication and transport. However, the building blocks composing membranes are not directly encoded by the genetic material; rather, they are synthesized through energetically expensive and complex metabolic pathways. In the quest for a SynCell, this project proposes to strip down complexity and swap lipids for peptide-based building blocks that are directly encoded by genetic material, that are programmable, and that can form interactions ranging from strong to weak, and specific to nonspecific. The plan is underpinned by the discovery of a variety of protein-based subcellular compartments in nature (bacterial microcompartments, phase separated droplets), and by the bottom-up peptide design of new membrane-forming building blocks with tunable properties and stimulus-responsive assembly/disassembly.

The long term goal of the ProteoCell project is to generate a completely programmable, self-sufficient cell that can replicate, generate its own energy, and assemble into multicellular structures, while offering completely new properties (robustness, catalytic functions, permeability) that are valuable to biochemical production and creation of synergistic cellular communities. Towards this goal, this project will:

Aim 1: Create a modular toolbox for compartment formation and encapsulation using polypeptides by generating composable, multi-compartment ProteoCells with a protein/peptide boundary using principles of macromolecular design, synthesis, hierarchical assembly, and phase separation. The ProteoCell will enclose its own genetic material and synthesize all its components via cell-free transcription-translation.

Aim 2: Create modules for compartmentalized biosynthesis within the ProteoCell by developing programmable, intracellular organelles spanning different sizes, morphologies, and permeabilities; demonstrating assembly within the ProteoCell cytoplasm of two types of rudimentary organelles; and demonstrating function for the production of reduced carbon compounds.

Aim 3: Create higher-order organization of ProteoCells to enable by-design construction of multicellular structures by functionalizing the exterior of ProteoCells to enable interaction with identical or different ProteoCells into tunable structures with long-term potential to create cell communities.

Aim 4: Contribute to responsible research and innovation of bottom-up synthetic biology and ProteoCells by assessing social and ethical implications of emerging public risk and benefit perceptions and integrating these findings into the scientific research enterprise and its outreach and education activities.

Intellectual merit. The proposed project will establish a minimalist SynCell, as well as portable parts and devices for applications in synthetic biology. The project will deliver a cell that is entirely programmable through its genetic material, in which parts can be swapped easily (e.g. natural components vs. designed versions), and that can be interfaced with other cells to form multicellular elements or tissues. The proposed ProteoCells address fundamental Rules of Life questions surrounding the biological archetypes for organization and compartmentalization e.g., are lipid assemblies required for a functional cell? Can life be simplified to three building blocks, down from the canonical four? The project will also yield an understanding of the societal impact of generating an artificial cell, including explicit assessment of risk perception. This feedback will inform current and future work on SynCells.

Broader impacts. The multidisciplinary team plans to integrate findings on public perceptions within all education and outreach activities. The concept of a SynCell offers a unique opportunity to galvanize the enthusiasm of K-12 students, undergraduates, and graduate students, and to engage the public in a conversation on the question What is a cell?. The plan is to offer a strategic set of education and outreach activities that will enrich undergraduate education through training of faculty at PUIs and community colleges, outreach to K-12 audiences at a crucial point in their scientific training in biological sciences, and engage in two-way dialogue with wider publics about the potentially transformative science and steps to ensure its responsible development.
StatusActive
Effective start/end date9/15/198/31/22

Funding

  • National Science Foundation (NSF): $700,662.00

Fingerprint

permeability
biology
education
community
organization
interaction
science
stimulus
conversation
student
dialogue
graduate
innovation
energy
communication