INDUCIBLE MOSAICISM

Smart overview of the Invention

The patent application outlines a novel approach for generating multiple unique genetic edits within the seeds of plants. This method involves introducing specific nucleic acids into plant cells or tissues. These nucleic acids encode a DNA modification enzyme, potentially accompanied by guide RNA (gRNA), and an inducible system sequence. This inducible system is activated at a desired growth stage, facilitating the expression or nuclear translocation of the DNA modification enzyme, leading to diverse genetic edits in floral primordia or reproductive organs.

Field of Application

Focusing on plant biotechnology, this invention pertains to gene editing in plants, specifically through inducible systems that enable multiple genetic modifications in plant progeny. The goal is to enhance the efficiency of obtaining diverse genetic edits during specific stages of a plant's life cycle, thereby improving crop quality and yield through innovative biotechnological methods.

Background and Challenges

Gene editing is pivotal for crop improvement but poses challenges due to inefficiencies in achieving certain edits, such as allele replacements or large deletions. Traditional methods like CRISPR or TALENs face difficulties with specificity and efficiency, especially in plants where homologous recombination is less favored. This invention addresses these challenges by providing a cost-effective and efficient method to produce diverse allelic variations and overcome the bottlenecks in current gene editing practices.

Methodology

The technique involves an expression cassette introduced into plant cells, comprising a nucleic acid encoding a DNA modification enzyme and an inducible factor linked to it. This factor can be triggered by various chemicals, including antibiotics, metals, steroids, hormones, alcohols, or aldehydes. The inducible factor may act as a transcription or translocation effector, facilitating targeted gene edits at specific developmental stages. Various effector systems like alcA/alcR or LhG4/pOp are utilized to achieve precise control over gene editing processes.

Applications and Embodiments

The invention supports a wide range of DNA modification enzymes such as CRISPR nucleases and others like meganucleases or TALENs. It allows for diverse genetic modifications including indels, nucleotide substitutions, allele replacements, and more. The method is applicable to various plant species including dicotyledonous plants like Arabidopsis and soybean. The system promises to significantly enhance the efficiency of producing genetically diverse plant progenies.