# Engineering Cytoskeletal Motors

> **NIH NIH R01** · STANFORD UNIVERSITY · 2021 · $319,800

## Abstract

SUMMARY
 Diverse cytoskeletal motors perform essential cellular functions including spindle assembly, nuclear
positioning, and polarized transport of mRNA, proteins, and membranous cargos along microtubules and actin
filaments. Engineering biomolecular motors with tunable and dynamically controllable properties can provide
(1) rigorous tests of models relating molecular structures to mechanical functions, (2) novel tools for selective
perturbation of mechanical processes inside living cells, and (3) optimized components for complex tasks such
as molecular sorting and directed assembly in vitro. This project seeks to develop and characterize a
comprehensive set of modified cytoskeletal motors with defined properties — including speed, direction, and
force generation — than can be controlled using external cues such as light. A modular protein engineering
approach will be applied to both actin-based and microtubule-based transport. During successive design
cycles, chimeric motors will be constructed based on structural models, and then functionally characterized
using gliding filament assays, single fluorophore imaging, gold nanoparticle tracking, and optical trapping.
Complementary structural characterization using cryoelectron microscopy will be used to compare the
experimental conformations of filament-bound motors to the original structural designs, and to yield new
insights into class-specific structure-function relationships. Finally, pilot studies will be conducted to test the
function of engineered motors inside living cells.
 The specific aims of this project are (1) to create diverse myosin motors that exploit dynamic changes
in lever arm structure in order to shift gears — speed up, slow down, or change directions — when exposed to
blue light; (2) to develop diverse microtubule-based motors with artificial lever arms, including light-activated
gearshifts, by exploiting a mechanistic analogy between myosins and class-14 kinesins, and (3) to create
processive multimeric assemblies of controllable engineered myosins and kinesins, and characterize their
force-generating properties. If successful, this work will dramatically expand the potential applications of
engineered molecular motors, and provide unprecedented control over nanoscale motion. Genetically encoded
light- responsive motors will expand the optogenetics toolkit, complementing precise perturbations of ion
channels and intracellular signaling with spatiotemporal control of cytoskeletal transport and contractility.
Optogenetic control of bidirectional transport will enable dynamic relocalization of biomolecules and organelles;
highly processive and controllable motors will have potential applications in gene and drug delivery; and
controllable motors may be used to sort, shuttle, and concentrate analytes in microfabricated diagnostic
devices.

## Key facts

- **NIH application ID:** 10238890
- **Project number:** 5R01GM114627-04
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Zev Bryant
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $319,800
- **Award type:** 5
- **Project period:** 2018-09-10 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10238890

## Citation

> US National Institutes of Health, RePORTER application 10238890, Engineering Cytoskeletal Motors (5R01GM114627-04). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10238890. Licensed CC0.

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