Genetically Encoded Chlorite Dismutase for Cellular Oxygen Control
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Solution Overview
Problem
Current methods lack the ability to manipulate intracellular oxygen levels in living cells with high spatiotemporal precision and control, limiting the ability to address oxygen-related health issues and research applications.
Innovation Solution
Expression of chlorite dismutase (Cld) enzymes in eukaryotic cells, optionally with a chlorite transporter, to convert chlorite into oxygen and chloride, enabling localized and controlled oxygen production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to manipulate oxygen in living cells, then oxygen levels can be adjusted, but the ability to achieve high spatiotemporal precision and control is limited
Solution Approach 1:
The invention divides the oxygen manipulation function into separate modular components: a chlorite transporter protein that controls substrate uptake and a chlorite dismutase enzyme that catalyzes oxygen production. This segmentation allows independent optimization and precise spatial-temporal control of each function, resolving the contradiction between precision and versatility.
Solution Approach 2:
The invention introduces chlorite as an intermediary substrate that is non-toxic and can be transported into cells by the engineered transporter. This intermediary enables controlled oxygen generation without directly introducing toxic oxygen sources, achieving precise spatiotemporal control while maintaining cell viability.
2Quantity of substance
If oxygen is introduced into living cells, then oxygen availability increases, but cell viability may be compromised due to oxidative stress
Solution Approach 1:
The invention converts the potentially harmful effect of oxygen toxicity into a beneficial outcome by using chlorite dismutase to catalyze the conversion of chlorite (a stable, non-toxic substrate) into oxygen and chloride. This controlled enzymatic conversion generates oxygen locally without introducing direct oxidative stress, maintaining cell viability while increasing oxygen availability.
Solution Approach 2:
The engineered cellular system uses its own metabolic machinery - the expressed chlorite transporter and chlorite dismutase enzymes - to autonomously generate oxygen within the cell. This self-service approach allows the cell to produce its own oxygen supply in a controlled manner, avoiding external oxygen introduction that could cause oxidative damage.
3Productivity
If chlorite dismutase is expressed in eukaryotic cells, then oxygen can be generated, but the complexity of the genetic system increases
Solution Approach 1:
The invention uses a universal approach by selecting chlorite dismutase, an enzyme already known to catalyze oxygen production from chlorite. This pre-characterized enzyme can be expressed in various eukaryotic cell types using standard molecular biology techniques, achieving oxygen production without requiring development of entirely new complex systems.
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
Provides a genetically encoded system for generating oxygen in living cells, allowing for precise spatiotemporal control and maintaining cell viability, with potential applications in research and medical settings.
Implementation Method 1
an enzyme that converts chlorite into oxygen and chloride. This enzyme is abbreviated Cld and referred to as chlorite dismutase, chlorite O2-lyase, chlorite:O2 lyase, chlorite lyase, and chlorite oxidoreductase
Implementation Method 2
converts chlorite into oxygen and chloride
Implementation Method 3
a chlorite transporter, e.g., an exogenous chlorite transporter. In some embodiments, the chlorite transporter is a sodium iodide symporter (NIS)
Data Source
AI summary
Described herein are compositions and methods for generating oxygen in living eukaryotic cells, e.g., animal cells, by expressing a Cld enzyme (i.e., chlorite dismutase, chlorite O2-lyase, chlorite:O2 lyase), optionally in combination with a transporter, in the cells.