# Intracompartmental 3D Printing of Enzymatically Active Organelle Mimics

## TL;DR

Researchers created 3D-printed subcellular compartments in artificial cells that can localize and perform enzymatic reactions.

## Contribution

A method for intracompartmental 3D printing of enzymatically active organelle mimics in artificial cells.

## Key findings

- 3D-printed subcellular compartments were successfully created within coacervate-based artificial cells.
- The compartments could uptake and concentrate His6-tagged proteins despite increased viscosity.
- Enzymatic reactions were localized within specific regions of the artificial cells.

## Abstract

Introducing subcellular structures in artificial cells
is a key
step in mimicking the structure and role of organelles, which are
instrumental in compartmentalizing cellular reaction networks. Despite
the variety of strategies to include subcellular features within artificial
cell models, achieving spatial and morphological control over these
compartments remains challenging. In this study, we engineered 3D-printed
subcellular compartments within terpolymer-stabilized coacervate-based
artificial cells. Coacervate-forming charged polymers were functionalized
with methacrylate moieties, enabling the fabrication of a variety
of architectures within droplets through photoinitiated radical polymerization.
The addition of a Ni-NTA functional methacrylate monomer to the coacervates
led to its sequestration upon polymerization in these subcellular
regions. As a result, the compartments were able to uptake and concentrate
His6-tagged mTurquoise and β-galactosidase protein
cargo molecules, despite the increase in viscosity that was induced
upon polymerization. Following this affinity-based interaction approach,
we demonstrated the region-specific localization of an enzymatic reaction
within the artificial cells.