# New Biosensor Tools for Diaphanous Related Formin MD1A and 2, Useful for Multiplex Imaging

> **NIH NIH R01** · ALBERT EINSTEIN COLLEGE OF MEDICINE · 2020 · $22,329

## Abstract

Abstract/Summary
 Actin cytoskeleton rearrangement, an important determinant of both cellular motility and the maintenance
of cellular structure-function, is regulated by members of the Rho family of small GTPases. Rho family GTPase–
regulated proteins that catalyze actin fiber polymerization include those of the Wiskott Aldrich family (WASp/
WAVE) proteins acting on their downstream Arp2/3-complex, and formins including the Diaphanous-related
formin (DRF) subclass. How DRF activity is controlled is poorly understood compared with WASp/WAVE
proteins, however, particularly in relation to cellular functions driven by DRFs that are activated by Rho GTPases.
Our primary objective therefore, is to develop new fluorescent biosensor imaging tools for studying the DRFs
mammalian Diaphanous formin (mDia)1 and 2, for the first time. Our second objective is to extend these new
mDia biosensors and our Rho family GTPase biosensors to multiplex-imaging compatible format through the use
of our new near-infrared (NIR) fluorescent proteins (FPs) ideally suited for Förster resonance energy transfer
(FRET) applications. The new NIR FRET biosensors developed herein, will for the first time enable simultaneous
multiplex imaging and/or perturbation of upstream Rho GTPases and monitoring of mDia1 and 2 activities in
the same cell; the new NIR FRET modality will be optimized for simultaneous imaging using traditional cyan
fluorescent protein (CFP)–yellow fluorescent protein (YFP) FRET or combination with optogenetic tools that
require blue-green light. In summary, our proposed new biosensor technology for mDia1 and 2 will enable for
the first time direct, simultaneous monitoring and/or perturbation of Rho GTPase and mDia activities, allowing
determination of the specificity of Rho GTPase–initiated signalling dynamics regarding the functions of mDia1
and 2 in living cells. The approaches pursued in this proposal should significantly advance the current state of
the art in technology for studying RhoGTPase–mDia biology.

## Key facts

- **NIH application ID:** 10003872
- **Project number:** 3R01GM132098-01S1
- **Recipient organization:** ALBERT EINSTEIN COLLEGE OF MEDICINE
- **Principal Investigator:** Louis Hodgson
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $22,329
- **Award type:** 3
- **Project period:** 2019-07-01 → 2021-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10003872

## Citation

> US National Institutes of Health, RePORTER application 10003872, New Biosensor Tools for Diaphanous Related Formin MD1A and 2, Useful for Multiplex Imaging (3R01GM132098-01S1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10003872. Licensed CC0.

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