Nucleic Acid Constructs for In-Planta Fruit Tree Genome Editing
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Solution Overview
Problem
Conventional cross-breeding for developing new fruit tree varieties is time-consuming and inefficient, and existing genome editing methods in fruit trees are laborious and limited to research due to the need for callus formation and tissue culture, resulting in non-commercial transgenic plants.
Innovation Solution
An in-planta genome editing method using a nucleic acid construct system that includes genome editing agents and developmental regulators, combined with Agrobacterium delivery and T-DNA integration prevention, to produce transgene-free plants directly in soil-grown fruit trees.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional cross-breeding is used to develop new fruit tree varieties, then genetic diversity and trait combination are achieved, but the process is time-consuming and laborious
Solution Approach 1:
The patent replaces the mechanical biological process of cross-breeding with a molecular biology approach using CRISPR-Cas9 genome editing. Instead of physically crossing plants and waiting for generations to pass, the invention directly edits the genome of somatic cells to introduce desired traits, dramatically reducing the time required from decades to months or years.
Solution Approach 2:
The invention changes the fundamental parameter of trait introduction from indirect (through cross-breeding and selection) to direct (through genome editing). By using CRISPR-Cas9 to create precise mutations in target genes, the patent enables rapid introduction of specific traits without the lengthy process of traditional breeding.
2Manufacturing precision
If CRISPR/Cas9 genome editing is applied to fruit trees using existing methods, then precise trait modification is achieved, but the process requires callus formation and tissue culture resulting in non-commercial transgenic plants
Solution Approach 1:
The patent extracts and eliminates the complex tissue culture and callus formation steps from the genome editing process. By developing a method that delivers CRISPR-Cas9 components directly to somatic cells in living plants (in-planta editing), the invention removes the need for in vitro regeneration, simplifying the process and enabling production of non-transgenic edited plants suitable for commerce.
Solution Approach 2:
The invention enables the plant itself to serve as the site of genome editing by delivering CRISPR-Cas9 components directly to somatic cells in the living plant. This self-service approach eliminates the need for external tissue culture facilities and complex regeneration protocols, allowing the plant to regenerate naturally after editing.
3Reliability
If tissue culture is used for genome editing in fruit trees, then genome editing can be performed, but the process is cost-ineffective and limited to research
Solution Approach 1:
The patent replaces the costly and complex tissue culture system with a direct in-planta editing approach. By delivering CRISPR-Cas9 components directly to somatic cells in living plants through methods such as agrobacterium-mediated transformation or particle bombardment, the invention eliminates the need for expensive tissue culture facilities and reduces operational costs while maintaining editing reliability.
4Adaptability or versatility
If developmental regulators are introduced to induce meristem formation, then de novo edited meristem induction is achieved, but the process is not yet tested in fruit trees
Solution Approach 1:
The patent applies the meristem induction strategy developed in model plants to fruit trees, demonstrating the universal applicability of the approach. By introducing developmental regulator genes (such as WUS, STM, Ipt, and PLT5) alongside CRISPR-Cas9 components, the invention successfully induces meristem formation and edited shoot development in fruit trees, validating the method across different plant species.
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 method significantly improves de novo shoot induction and regeneration efficiency, enabling the production of homozygous/bi-allelic genome-edited plants and transgene-free progeny, accelerating the development of new cultivars.
Implementation Method 1
combined with Agrobacterium delivery and T-DNA integration prevention
Data Source
AI summary
A nucleic acid construct or construct system for genome editing is provided. Also provided are non-transgenic plants having been edited accordingly and methods of producing such plants.