Transient Morphogenic Regulators for Transgene-Free Plant Regeneration
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Solution Overview
Problem
Conventional breeding and genetic engineering methods struggle to efficiently produce transgene-free plants, particularly in vegetative propagated crops, due to their heterozygotic nature and multiploidy, leading to technical challenges in segregating away undesired foreign DNA and random mutations.
Innovation Solution
The use of heterologous morphogenic regulators introduced into plant cells through polynucleotides or polypeptides to enhance regeneration frequency and stimulate the formation of transgene-free gene-edited plants, utilizing gene-editing systems for precise nucleic acid modification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional breeding methods are used to segregate away transgenes in vegetative propagated crops, then transgene-free plants can be produced, but the process requires multiple rounds of crossing which is technically challenging and economically undesirable
Solution Approach 1:
The patent extracts and eliminates foreign DNA and transgenes from the plant genome using targeted genome editing tools (such as CRISPR-Cas9, TALEN, or zinc finger nucleases) that specifically recognize and cut sequences containing transgenes, thereby removing them without requiring complex multi-generational breeding procedures
Solution Approach 2:
The patent replaces the mechanical/conventional breeding approach (repeated crossing and selection) with a molecular biology-based genome editing system that directly modifies the DNA sequence to eliminate transgenes, achieving the same goal through a more direct and less complex mechanism
2Reliability
If recurrent crossing is performed to segregate away transgenes in heterozygotic and multiploid plants, then transgene elimination can be achieved, but it requires multiple rounds of crossing which causes genetic drift and loss of beneficial alleles
Solution Approach 1:
The patent extracts and removes only the unwanted transgenes and foreign DNA through targeted genome editing, while leaving the beneficial alleles and genetic background intact, thereby eliminating the need for repeated crossing that would otherwise cause loss of beneficial genetic information
Solution Approach 2:
The patent applies localized genome editing at specific positions in the genome where transgenes are located, rather than performing global genetic reshuffling through repeated crossing, thereby maintaining the overall genetic integrity and beneficial alleles of the plant while removing only the problematic transgene sequences
3Manufacturing precision
If conventional genetic engineering methods are used to introduce targeted genetic modification, then gene editing can be achieved, but identifying and removing random mutations caused by integration of foreign DNAs remains technically challenging
Solution Approach 1:
The patent incorporates molecular markers, fluorescent tags, or other detectable elements that provide feedback signals to identify and track the integration sites of foreign DNAs and transgenes, enabling precise detection and removal of random mutations through targeted genome editing without requiring complex analysis procedures
Data Source
AI summary
Methods and compositions for generating transgene-free plants are described. In particular, the transgene-free plants are regenerated by transient expression of morphogenic regulators in a plant cell that is precisely edited using a gene-editing methodology. Also, provided are transformed plants, plant cell, tissues, and seeds in which a target gene is edited, but non-transgenic.