# Multiplex single-cell analysis of intracellular protein signaling dynamics

> **NIH NIH R21** · UNIVERSITY OF CALIFORNIA RIVERSIDE · 2021 · $184,262

## Abstract

ABSTRACT
Recent developments on single-cell analytical methods have provided a clarifying view on cellular heterogeneity
and its associated mechanistic and therapeutic implications. Nevertheless, single-cell studies on intracellular
protein signaling activities can be confounded by the dynamic nature of signaling events. In addition, there is a
pressing need of developing non-genetic analytical methods for clinical samples. These requisites call for a
single-cell approach that can interrogate intracellular protein signaling dynamics while being compatible with
other downstream analytical methods. A recently developed prototype method has enabled dynamic probing of
intracellular protein signaling activities at single-cell resolution, without genetic modifications. The technology
was based on intracellularly delivered epitope-targeting peptide probes, a microwell-based single-cell chip and
high-speed imaging techniques. However, many obstacles exist that challenge the generalizability of this
approach and limit its biomedical application. In this proposal, the aim is to significantly advance this proof-of-
concept technology. Five specific goals are included: develop an automated screening protocol for improved
screening efficiencies, implement a unique bicyclic peptide library for higher biding affinities, employ cell
permeabilizers as a more generalizable delivery method, achieve simultaneous analysis of multiple signaling
proteins, and integrate this technology with other single-cell detection methods. This integrated multiplex pipeline
will provide an enabling technology that promises a more in-depth understanding of the interplay among protein
signaling activities, protein expression levels and metabolism in biological processes.

## Key facts

- **NIH application ID:** 10152653
- **Project number:** 5R21EB025393-03
- **Recipient organization:** UNIVERSITY OF CALIFORNIA RIVERSIDE
- **Principal Investigator:** Min Xue
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $184,262
- **Award type:** 5
- **Project period:** 2019-08-07 → 2023-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10152653

## Citation

> US National Institutes of Health, RePORTER application 10152653, Multiplex single-cell analysis of intracellular protein signaling dynamics (5R21EB025393-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10152653. Licensed CC0.

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