Cannabinoid Biosynthesis Control Using CRISPR Transcription Regulation
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Solution Overview
Problem
Current methods for regulating cannabinoid production in plants often require permanent genetic modifications and lack the ability to selectively control the production of specific cannabinoids without altering the plant's genetic material.
Innovation Solution
A method utilizing CRISPR-Cas technology to introduce nucleotide sequences that regulate the endogenous transcriptional machinery of plants, allowing for the selective increase or decrease of cannabinoid production by targeting specific genes in the biosynthetic pathway through guide RNAs, inactive Cas nucleases, and transcriptional activators or repressors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If permanent genetic modification methods are used to regulate cannabinoid production, then the ability to selectively control specific cannabinoid production is improved, but the complexity of permanently altering plant genetic material increases
Solution Approach 1:
The patent uses CRISPR-Cas9 system as an intermediary tool to regulate cannabinoid production without permanent genetic modification. The guide RNA sequences act as mediators that direct the Cas nuclease to specific target genes in the cannabinoid biosynthetic pathway, enabling selective control of cannabinoid production while avoiding the complexity of permanent genetic alteration. This temporary, reversible approach allows precise regulation through molecular intermediaries rather than direct genomic changes.
2Ease of operation
If simple genetic techniques like cross fertilization are used to change plant properties, then the ease of operation is improved, but the precision of regulating specific cannabinoid production decreases
Solution Approach 1:
The patent replaces traditional mechanical breeding methods (cross-fertilization) with a molecular-level approach using CRISPR-Cas9 technology. Instead of relying on random genetic recombination through pollination, the system uses guide RNA sequences to precisely target and regulate specific genes encoding enzymes in the cannabinoid biosynthetic pathway. This substitution of mechanical breeding with molecular precision editing enables exact control over which cannabinoids are produced and in what quantities, achieving high manufacturing precision while maintaining operational simplicity through standardized molecular protocols.
3Productivity
If overexpression of CBDAS gene and suppression of THCAS gene through RNA interference is used, then the productivity of CBD is improved, but the loss of THC production occurs
Solution Approach 1:
The patent implements dynamic, reversible regulation of cannabinoid production using inducible CRISPR-Cas9 systems. Instead of permanent overexpression or suppression, the system allows flexible adjustment of guide RNA expression levels and Cas nuclease activity to dynamically control the balance between CBD and THC production. This dynamic approach enables producers to optimize cannabinoid profiles based on market demands, temporarily enhancing CBD productivity when needed while preserving the ability to restore THC production by adjusting or removing the CRISPR components, thus avoiding permanent loss of substance.
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 controlled and selective enhancement or reduction of cannabinoid levels in plants, achieving increases or decreases of up to 600-fold without creating genetically modified organisms, providing flexibility and efficiency in producing desired cannabinoid profiles.
Implementation Method 1
the first nucleotide sequence comprising at least one guide RNA sequence directed towards a plant gene that encodes a first protein in the biosynthetic pathway of the least one cannabinoid
Implementation Method 2
at least one sequence encoding a transcriptional activator or a repressor protein, and wherein, when the first nucleotide sequence encodes a transcriptional activator
Data Source
Figure 1A~1C
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AI summary
Disclosed is a method of regulating production of at least one cannabinoid in plants, through the introduction of nucleotide sequences into the plant that result in one or more of: overexpression of a gene of interest to increase production of a cannabinoid of interest; enhanced production of the protein of interest by CRISPR-TAD; repression of production of competitive proteins by CRISPR-REP to promote in an indirect manner the production of the cannabinoid of interest. Also disclosed are nucleic acids for the genetic modification and plants, seeds or plant parts that result from the modification.