# Acquisition of a Confocal  Fluorescence Microscope to Image Synthetic Biomolecular Condensates

> **NIH NIH R35** · DUKE UNIVERSITY · 2022 · $197,907

## Abstract

Abstract
The proposed work in this MIRA application leverages my long-standing interest and expertise in the design of
genetically encoded stimulus-responsive peptide polymers. My group pioneered the development of
recombinant elastin-like polypeptides (ELPs) that exhibit lower critical solution temperature (LCST) phase
behavior. We have also, in parallel, pioneered the development of high-throughput methods for the assembly
of highly repetitive genes that we used to create the largest extant library of recombinant peptide polymers.
Characterization of their aqueous phase behavior led to the discovery of sequence heuristics that can be used
for the de novo used design of peptide polymers that exhibit LCST phase behavior and a class of resilin-like
polypeptides (RLPs) that exhibit the converse — upper critical solution phase transition (UCST) phase
behavior. Building upon this work, we will explore two new areas in this proposal. First, we will investigate how
we can recapitulate the hierarchical structure and properties exhibited by biological materials by the design of
partially ordered polymers (POPs) —that consist of disordered polypeptides embedded with a periodically
recurring secondary structure motif— that exhibit temperature triggered hierarchical self-assembly into
macroscopic materials that mimic the in vivo organization of structural proteins like elastin networks. We will
carry out a systematic exploration of the design of new POPs, to verify that the combination of order and
disorder at the chain segment level is a new and robust design principle that will yield materials with
hierarchical selfassembly across many length scales. Second, we will develop a new line of investigation on
genetically encoded biohybrid polymers via post-translational modifications (PTMs) that precisely combine
peptide and non-peptide components to create biomaterials that exhibit triggered, hierarchical self-assembly
into macroscopic materials. In this aim, we will expand upon our initial work on in vivo myristoylation of ELPs
and UCST exhibiting RLPs to investigate if we can convert structure-directing peptides into myristoylation
substrates, to create myristoylated polypeptides where the myristoylated segment can direct hierarchical self-
assembly of the entire construct. We will also investigate modification of ELPs and RLPs with cholesterol that
has the potential to direct self-assembly, and phosphorylation, which will provide a unique trigger of self-
assembly. Much remains to be done in both areas, as our preliminary foray into these new areas only hint at
the enormous possibilities in the molecular design of new biomaterials enabled by these approaches. The work
we propose herein promises to yield new biomaterials with interesting structures and properties with a host of
applications in biotechnology and medicine.

## Key facts

- **NIH application ID:** 10581200
- **Project number:** 3R35GM127042-05S1
- **Recipient organization:** DUKE UNIVERSITY
- **Principal Investigator:** Ashutosh Chilkoti
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $197,907
- **Award type:** 3
- **Project period:** 2018-09-01 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10581200

## Citation

> US National Institutes of Health, RePORTER application 10581200, Acquisition of a Confocal  Fluorescence Microscope to Image Synthetic Biomolecular Condensates (3R35GM127042-05S1). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10581200. Licensed CC0.

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