# Bottom-up, high-throughput prototyping of extracellular vesicle mimetics using cell-free synthetic biology

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA AT DAVIS · 2024 · $561,799

## Abstract

PROJECT SUMMARY
Bottom-up, high-throughput prototyping of extracellular vesicle mimetics using cell-free synthetic biology
Cells secrete extracellular vesicles (EVs) that function as primary messengers of intercellular communication
and are studied as promising drug-delivery vehicles and therapeutics. However, the clinical application of native
EVs has been hindered by their low production yield, impurity, and inherent heterogeneity. Native EVs contain
many biologically active components, such as RNAs and proteins, spread out over numerous subpopulations.
This biological complexity is both the strength and the Achilles’ heel of native EVs. While various features of this
complexity enable the beneficial therapeutic effects of EVs, it is not clear which plays a dominant role. However,
the complex set of proteins and RNAs results in heterogeneous EVs that are challenging to study and use as a
standardized treatment. Therefore, separating out and defining the critical biomolecular features from the overall
heterogeneous set will allow us to perform quality control of EVs and to reproducibly produce or study EVs. A
major bottleneck in finding the critical molecular parts of EVs is the lack of high-throughput methods. To
overcome this difficulty, our team will create a synthetic biology-based, cell-free high-throughput discovery
platform. The platform will be able to synthesize EV mimetics using a cell-free synthesis approach (Aim 1),
coupled with high-throughput examination of EV mimetic potency in vitro (Aim 2). Select EV mimetics will also
be investigated using an in vivo model system of neuroprotection and immune modulation (Aim 3). Throughout
the study, we will use native mesenchymal stem/stromal cell EVs and neurological diseases as our model system
to evaluate the platform. Our work will enable the high-throughput study of EVs for any disease and biological
questions of interest. In addition, we will unveil new insights into EVs that address key debated topics in the EV
field.

## Key facts

- **NIH application ID:** 10794334
- **Project number:** 5R01EB034279-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA AT DAVIS
- **Principal Investigator:** Randy Carney
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $561,799
- **Award type:** 5
- **Project period:** 2023-03-01 → 2027-02-27

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10794334

## Citation

> US National Institutes of Health, RePORTER application 10794334, Bottom-up, high-throughput prototyping of extracellular vesicle mimetics using cell-free synthetic biology (5R01EB034279-02). Retrieved via AI Analytics 2026-06-12 from https://api.ai-analytics.org/grant/nih/10794334. Licensed CC0.

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