Modified Luciferase Mutations for Brighter Autoluminescence Imaging
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Solution Overview
Problem
The luminescence intensity of autoluminescence systems derived from luminescent mushrooms limits the observation of physiological phenomena within or between cells, necessitating an improvement in luminescence intensity for effective molecular imaging.
Innovation Solution
A modified luciferase is introduced with mutations at specific amino acid positions (arginine or lysine at position 26 and valine at position 161) to enhance luminescence intensity, utilizing a luminescent mushroom-derived luciferase with improved catalytic efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If a luminescent mushroom-derived luciferase system is used for autoluminescence imaging, then the system can perform in vivo observation without adding external luciferin, but the luminescence intensity is insufficient for effective observation of physiological phenomena
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid mutations at positions 26 (arginine or lysine) and 161 (valine) in the luciferase protein sequence. These mutations alter the catalytic parameters of the enzyme, enhancing its activity and consequently increasing the luminescence intensity produced during the luciferin oxidation reaction, while maintaining the autonomous imaging capability of the system
2Reliability
If wild-type luciferase is used, then the protein structure is stable and well-characterized, but the luminescence intensity is insufficient for observing physiological phenomena within cells
Solution Approach 1:
The patent applies local quality by making targeted mutations at specific positions (26 and 161) in the luciferase protein sequence rather than completely redesigning the protein. This approach maintains the overall stable structure and well-characterized properties of wild-type luciferase while locally improving the catalytic efficiency at specific sites to enhance luminescence intensity
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
The modified luciferase exhibits enhanced luminescence intensity, potentially improving the observation of physiological phenomena by increasing light emission, thereby enhancing molecular imaging capabilities.
Implementation Method 1
Chemiluminescence imaging uses a luminescence phenomenon (luciferin-luciferase reaction) caused by an oxidation reaction of a luminescent substrate (luciferin) catalyzed by a chemiluminescent protein (luciferase)
Data Source
AI summary
The present disclosure, in one aspect, provides a modified luciferase with improved luminescence intensity, and use thereof. The present disclosure relates to a modified luciferase derived from a luminescent mushroom or a functional fragment thereof, including a mutation introduced into arginine or lysine corresponding to position 26 and/or a mutation introduced into valine corresponding to position 161 of amino acid sequence shown in SEQ ID NO: 1, and having enhanced luminescence compared with a luciferase before the mutation is introduced. The present disclosure also relates to an isolated nucleic acid encoding the modified luciferase or functional fragment thereof of the present disclosure.