Rosaceae TFL Gene Editing for Early Flowering and Shorter Juvenile Phase
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Solution Overview
Problem
Genetic gain through breeding in Rosaceae plants is limited by environmental controls that trigger flowering and a long juvenile phase, which hinders efficient plant production.
Innovation Solution
Modifying the TERMINAL FLOWER (TFL) gene in Rosaceae plants using CRISPR-Cas editing systems to introduce mutations, reducing the dependency on environmental triggers for flowering and enhancing flowering time and yield characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If natural breeding methods are used in Rosaceae plants, then genetic diversity is maintained, but flowering time is controlled by environmental factors and juvenile phase is prolonged
Solution Approach 1:
The patent applies parameter changes by modifying the TFL1 gene through CRISPR-Cas9 editing to alter its expression levels and function. This genetic parameter modification directly reduces the juvenile phase duration and eliminates environmental dependency for flowering initiation, transforming the plant's developmental timing parameters without requiring prolonged environmental conditioning
Solution Approach 2:
The patent replaces the natural mechanical/environmental triggering system for flowering with a genetically engineered system. Instead of relying on environmental cues (light, temperature) to trigger flowering after a long juvenile phase, the modified TFL1 gene creates an intrinsic genetic mechanism that initiates flowering autonomously, substituting environmental control with genetic control
2Reliability
If natural variation in flowering time genes is selected, then environmental dependency is reduced, but such variation is rare and limited to a few species
Solution Approach 1:
The patent replaces traditional mechanical breeding methods (selection and crossing) with a molecular biology-based CRISPR-Cas9 gene editing system. This substitution allows direct modification of the TFL1 gene in any Rosaceae species, eliminating the need to wait for rare natural mutations and enabling efficient, targeted genetic improvement across multiple species
Solution Approach 2:
The patent enables precise parameter changes in the TFL1 gene through CRISPR-Cas9 editing, allowing researchers to modify specific nucleotides, delete portions of the gene, or adjust expression levels. This provides controlled and predictable changes in flowering time characteristics, unlike the random and rare natural variations available through traditional breeding
3Productivity
If TFL gene modification is performed using CRISPR-Cas systems, then flowering time and yield characteristics are improved, but transgene presence may complicate regulatory approval
Solution Approach 1:
The patent applies the discarding and recovering principle by designing the CRISPR-Cas9 system to be transient. The Cas9 protein and guide RNA are introduced temporarily to perform the gene edit, then discarded as they do not integrate into the plant genome. The resulting plants contain only the desired TFL1 modification without foreign transgenes, simplifying regulatory approval while maintaining improved productivity
Solution Approach 2:
The patent uses a transient expression system where CRISPR-Cas9 components are delivered temporarily (via Agrobacterium or protoplast transformation) rather than being permanently integrated. This substitutes permanent transgenic approaches with a transient editing approach, achieving the same genetic modification without leaving behind complex transgene structures that would complicate regulatory review
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 method results in Rosaceae plants with reduced time to flowering, extended flowering duration, and improved yield characteristics, overcoming the limitations of natural variation.
Implementation Method 1
The present invention provides methods for modifying the flowering time in Rosaceae plants by cleaving, in a site-specific manner, a target site within an endogenous TERMINAL FLOWER (TFL) gene in a plant cell using a CRISPR-Cas effector protein and a guide nucleic acid
Data Source
AI summary
This invention relates to compositions and methods for modifying TFL genes in Rosaceae plants to remove or reduce the dependency on environmental triggers for flowering, optionally producing plants having improved characteristics for breeding and production, optionally, including a reduced time to initiate flowering, a longer duration of flowering, and/or improved yield characteristics. The invention further relates to Rosaceae plants produced using the methods and compositions of the invention.