Bacteriophytochrome NIR Light-Activated Proteins

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Solution Overview

Problem

Current genetic and pharmaceutical approaches lack the spatiotemporal resolution and target specificity to accurately interrogate cellular functions in real time in vivo, limiting the ability to precisely activate or inactivate desired proteins in specific cells or tissues of live animals.

Innovation Solution

Development of genetically engineered far-red/NIR-light activated homodimeric proteins (NIRLAHPs) using photosensory modules of bacteriophytochromes, which can be activated or inactivated by far-red/NIR light, allowing for precise control of cellular processes with high spatial and temporal precision.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If natural photoactivated proteins (e.g., channelrhodopsins) are used for optogenetic approaches, then neurobiology insights are improved, but tissue penetration depth is limited to millimeter scale

Engineering Contradiction:
Improvespatiotemporal resolutionVSAvoidtissue penetration depth
Core Design Contradiction:
Measurement precisionVSLength of stationary object

Solution Approach 1:

The patent changes the spectral parameter of the photoreceptor proteins from visible-light absorbing (channelrhodopsins) to far-red/NIR-light absorbing (bacteriophytochromes). This parameter change in absorption wavelength enables deeper tissue penetration (centimeter scale) while maintaining optogenetic control capability, resolving the contradiction between measurement precision and tissue penetration depth.

Inventive Principle:
Principle #35Parameter changes

2Length of stationary object

If far-red/NIR light-activated proteins are used, then tissue penetration depth is improved to centimeter scale, but the ability to engineer proteins with desired output activities is limited

Engineering Contradiction:
Improvetissue penetration depthVSAvoidprotein output activity engineering
Core Design Contradiction:
Length of stationary objectVSAdaptability or versatility

Solution Approach 1:

The patent segments the photoreceptor system into modular components: a far-red/NIR light-sensitive bacteriophytochrome domain and separate output domains (enzymatic activities, binding specificities). This segmentation allows independent engineering of each module, enabling versatile protein output activities while maintaining deep tissue penetration capability through the bacteriophytochrome component.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent creates universal bacteriophytochrome-based photoactivated proteins that can perform multiple functions by combining the common bacteriophytochrome light-sensing module with various output domains. This universality enables the same light-activated platform to control diverse cellular processes (metabolism, signaling, apoptosis, hormone regulation) while maintaining centimeter-scale tissue penetration.

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Ease of operation

If visible light is used for protein activation, then protein activation capability is achieved, but light penetration into animal tissues is limited to millimeter scale

Engineering Contradiction:
Improveprotein activation capabilityVSAvoidlight penetration depth
Core Design Contradiction:
Ease of operationVSLength of stationary object

Solution Approach 1:

The patent changes the operational parameter from visible light activation to far-red/NIR light activation by using bacteriophytochromes instead of conventional photoreceptors. This parameter change in activation wavelength enables deep tissue penetration (centimeters) while preserving full protein activation capability, as bacteriophytochromes undergo robust conformational changes upon far-red/NIR light absorption that can be coupled to output domains.

Inventive Principle:
Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Enables the regulation of diverse cellular processes with high precision, including metabolic enzymes, signal transduction, cell apoptosis, and hormone regulation, offering potential applications in medical treatments such as cancer therapy and hormone deficiencies, as well as industrial uses like biofilm dispersion.

Implementation Method 1

Far-red/NIR light penetrates animal tissues much deeper (centimeter scale) than visible light (millimeter scale) absorbed by currently used photoreceptors

Methodology Applied
Scientific EffectLight absorption: Absorption (EM radiation)

Data Source

PatentUS10041057B2Near-infrared light-activated proteins
Publication Date: 2018.08.07 UNIVERSITY OF WYOMING

AI summary

Methods and constructs are provided for controlling processes in live animals, plants or microbes via genetically engineered near-infrared light-activated or light-inactivated proteins including chimeras including the photosensory modules of bacteriohytochromes and output modules that possess enzymatic activity and/or ability to bind to DNA, RNA, protein, or small molecules. DNA encoding these proteins are introduced as genes into live animals, plants or microbes, where their activities can be turned on by near-infrared light, controlled by the intensity of light, and turned off by near-infrared light of a different wavelength than the activating light. These proteins can regulate diverse cellular processes with high spatial and temporal precision, in a nontoxic manner, often using external light sources. For example, near-infrared light-activated proteins possessing nucleotidyl cyclase, protein kinase, protease, DNA-binding and RNA-binding activities are useful to control signal transduction, cell apoptosis, proliferation, adhesion, differentiation and other cell processes.