Dummy-Fluorescent Polypeptides Reduce Cellular Toxicity

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

In optogenetics, the expression of fluorescent polypeptides can lead to photo bleaching and toxicity in cells due to excitation by light, reducing the effectiveness of procedures and therapies.

Innovation Solution

Development of dummy-fluorescent polypeptides, such as mutated tdTomato, which are used in fusion proteins to reduce or eliminate fluorescence, thereby minimizing toxicity and enhancing the safety and efficacy of optogenetic assays and therapies.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If fluorescent polypeptides are expressed in cells for optogenetic procedures, then cell visualization and tracking are improved, but cellular toxicity and photo bleaching increase

Engineering Contradiction:

Improvecell visualizationVSAvoidcellular toxicity

Core Design Contradiction:

Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The patent applies parameter changes by modifying the fluorescent polypeptide's fluorescence intensity parameter. Dummy-fluorescent polypeptides are engineered to have dramatically reduced fluorescence emission (10-1000 fold reduction) while maintaining other functional properties, thereby eliminating toxicity associated with high fluorescence without sacrificing visualization capability

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses copying by creating dummy-fluorescent polypeptides that replicate the structural and functional properties of wild-type fluorescent polypeptides but with modified fluorescence characteristics. These dummy versions serve as functional copies that maintain protein-protein interactions and cellular localization while eliminating harmful fluorescence-related toxicity

Inventive Principle:

Principle #26Copying

2Loss of information

If fluorescent polypeptides are used in fusion proteins, then protein localization and expression tracking are improved, but photo bleaching and cellular damage increase

Engineering Contradiction:

Improveexpression trackingVSAvoidtherapy effectiveness

Core Design Contradiction:

Loss of informationVSDuration of action of stationary object

Solution Approach 1:

The patent changes the fluorescence intensity parameter of the polypeptide to create dummy-fluorescent versions that provide sufficient signal for tracking protein localization and expression while dramatically reducing photo bleaching and cellular damage, thereby extending the duration of therapeutic action

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses dummy-fluorescent polypeptides as intermediary components in fusion proteins that mediate between the need for expression tracking and the need to minimize cellular damage. These intermediaries provide the necessary tracking function with reduced harmful effects, enabling long-term therapeutic effectiveness

Inventive Principle:

Principle #24Intermediary (Mediator)

Data Source

PatentUS11278633B2Dummy-fluorescent protein fusions and methods of use

Publication Date: 2022.03.22 MASSACHUSETTS INST OF TECH

AI summary

The invention, in some aspects, relates to dummy-fluorescent (DF) polypeptide molecules and their encoding nucleic acid molecules and use of such molecules in fusion proteins and their encoding nucleic acid molecules. Compositions of the invention may be delivered to cells and subjects and used in methods to modulate electrical activity of cells in which they are expressed, and for treatment of diseases and conditions in subjects.