Tagged Bacteria Using CRISPR for GMO-Free Strain Tracking
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Solution Overview
Problem
There is a need for methods to easily tag microbial strains, particularly bacteria used as starter cultures in the fermented food/beverage industry, to identify their source and monitor their use, while avoiding regulatory issues associated with genetically modified organisms and addressing challenges such as contamination and bacteriophage resistance.
Innovation Solution
The method involves integrating a sequence from a bacteriophage into the CRISPR locus of bacteria, creating a unique tag that is not genetically modified, using CRISPR variants to identify the tagged bacteria by comparing nucleotide sequences with a database, and utilizing CRISPR spacers and repeats to confer bacteriophage resistance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional tagging methods using genetic modification are used, then identification capability is improved, but regulatory compliance and consumer acceptance deteriorate
Solution Approach 1:
The patent converts the harmful effect of bacteriophage contamination into a beneficial tagging mechanism. By allowing bacteriophages to naturally infect bacteria and integrate their genetic material into the CRISPR locus, the system creates unique identification tags without requiring artificial genetic modification. This natural infection process, which would normally be harmful, is harnessed to achieve the desired identification capability while avoiding regulatory issues associated with GMOs.
2Loss of information
If bacteriophage exposure is used for tagging, then unique identification is improved, but bacteriophage resistance is worsened
Solution Approach 1:
The patent applies preliminary action by exposing bacteria to bacteriophages during the tagging process before the bacteria are deployed in food products. This pre-exposure allows the bacteria to develop immunity through CRISPR-mediated adaptive immune response, where the bacteriophage genetic material is integrated into the CRISPR locus as a tag. The bacteria gain both identification capability and resistance to the specific bacteriophage strain used for tagging.
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
This approach allows for effective tracking and identification of bacterial strains without genetic modification, providing a robust means to monitor their use and resistance to bacteriophages, while maintaining their viability in food products.
Implementation Method 1
integrating a sequence from a bacteriophage into the CRISPR locus of bacteria, creating a unique tag
Implementation Method 2
utilizing CRISPR spacers and repeats to confer bacteriophage resistance
Data Source
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AI summary
The present invention provides methods for tagging and/or identifying microorganisms. In some preferred embodiments, the microorganisms are bacteria. In some particularly preferred embodiments, the bacteria are members of the genus Streptococcus, while in other embodiments, the bacteria are members of other genera. The present invention also provides microorganisms tagged using the methods set forth herein. In some preferred embodiments, the tagged microorganisms are bacteria. In some particularly preferred embodiments, the tagged bacteria are members of the genus Streptococcus, while in other embodiments, the tagged bacteria are members of other genera.