CRISPR Editing of Endogenous PUF Genes for Plant Yield
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Solution Overview
Problem
Current breeding methods for increasing plant yield have plateaued, and transgenic approaches to modify hormone-related pathways have been unsuccessful in achieving significant yield improvements due to the complexity of yield traits and the need for fine-tuning genetic modifications.
Innovation Solution
The development of compositions and methods involving CRISPR-Cas effector proteins and guide nucleic acids to introduce mutations in endogenous PUF genes in plants, which encode pumilio domain-containing polypeptides, with the aim of enhancing yield traits.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If transgenic approaches are used to modify hormone-related pathways, then yield improvement is attempted, but the approach fails to achieve significant yield improvements due to the complexity of yield traits and the need for fine-tuning
Solution Approach 1:
The patent changes the approach from transgenic over-expression to CRISPR-Cas9-mediated precise gene editing. This involves changing the modification strategy from adding foreign genes to precisely editing endogenous genes at specific nucleotide positions, enabling fine-tuning of hormone-related pathways to achieve reliable yield improvement
Solution Approach 2:
The patent replaces the mechanical transgenic approach (stable transformation of foreign DNA) with a molecular biology-based CRISPR-Cas9 system. This substitution enables more precise and controllable genetic modification, addressing the reliability issue by allowing targeted edits to specific genes involved in yield-related pathways
2Productivity
If multiple small-effect genes are assembled in a breeding program, then yield improvement is attempted, but research and development costs are substantially increased
Solution Approach 1:
The patent combines multiple beneficial mutations into a single CRISPR-Cas9 editing strategy. By targeting multiple genes simultaneously or sequentially with the same technology platform, the patent consolidates what would otherwise require multiple separate breeding programs, thereby reducing overall research and development resource requirements
Solution Approach 2:
The patent performs preliminary identification and validation of target genes and editing sites before large-scale breeding deployment. By pre-characterizing the effects of specific mutations in model systems or controlled environments, the patent reduces the resources needed for subsequent field trials and breeding programs
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 the targeted modification of PUF genes to improve yield traits in plants, potentially leading to increased seed size, seed oil content, and overall yield, while avoiding the limitations of transgenic approaches.
Implementation Method 1
a guide nucleic acid (e.g., gRNA, gDNA, crRNA, crDNA, sgRNA, sgDNA) comprising a spacer sequence with complementarity to an endogenous target gene encoding a PUF polypeptide
Data Source
AI summary
This invention relates to compositions and methods for modifying a PUF gene in plants, optionally to improve yield traits. The invention further relates to plants having increased improved yield traits produced using the methods and compositions of the invention.