CRISPR and RNAi Suppression of Flowering in Sugarcane
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Solution Overview
Problem
The challenge in sugarcane and energycane is to control flowering time effectively, as flowering leads to reduced sugar yield and extractability due to the cessation of vegetative growth and dehydration of stalk tissues.
Innovation Solution
The use of CRISPR constructs and RNAi systems to target and inhibit the expression of FT4, FT8, and FT10 genes, which are responsible for the expression of florigens in sugarcane and energycane, resulting in suppressed or delayed flowering.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If flowering is allowed to occur naturally in sugarcane and energycane, then reproductive development proceeds, but sugar yield and extractability decrease due to cessation of vegetative growth and stalk dehydration
Solution Approach 1:
The patent extracts and removes the harmful flowering function by targeting and disabling specific genes (FT4, FT8, FT10) that control flowering. Using CRISPR-Cas9 gene editing and RNA interference technologies, the invention selectively eliminates the flowering pathway while preserving all other plant functions, thereby preventing the cessation of vegetative growth and maintaining sugar production capacity.
Solution Approach 2:
The patent changes the genetic parameters of the sugarcane and energycane plants by modifying flowering-related genes. Through CRISPR-Cas9 editing and RNAi, the expression levels of FT4, FT8, and FT10 genes are altered or completely suppressed, fundamentally changing the plant's developmental parameters from natural flowering to continuous vegetative growth, thereby maintaining high sugar yield and extractability.
2Reliability
If flowering occurs in sugarcane and energycane, then reproductive development is achieved, but stalk density and sugar extractability are compromised due to tissue dehydration
Solution Approach 1:
The patent extracts and removes the harmful dehydration effect by eliminating the flowering trigger mechanism. By disabling FT4, FT8, and FT10 genes through CRISPR-Cas9 and RNAi, the invention prevents the signaling cascade that leads to reproductive development and associated stalk dehydration, thereby maintaining consistent sugar extractability without the harmful side effects of flowering.
3Productivity
If CRISPR constructs and RNAi systems are used to inhibit FT4, FT8, and FT10 genes, then flowering is suppressed or delayed by at least 50% to 100%, but genetic modification complexity increases
Solution Approach 1:
The patent uses intermediary molecular mechanisms to achieve flowering suppression. CRISPR-Cas9 serves as an intermediary gene-editing tool that introduces precise mutations in FT genes, while RNA interference acts as an intermediary that degrades target mRNA. These intermediary systems enable effective flowering suppression with controlled complexity, balancing genetic modification requirements with desired productivity outcomes.
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 achieves a significant suppression or delay of flowering by at least 50% to 100% or by several weeks to months, leading to increased sugar and biomass yield.
Implementation Method 1
The use of CRISPR constructs and RNAi systems to target and inhibit the expression of FT4, FT8, and FT10 genes
Implementation Method 2
The use of CRISPR constructs and RNAi systems to target and inhibit the expression of FT4, FT8, and FT10 genes
Data Source
AI summary
Described herein are compositions, including RNAi and/or CRISPR constructs, for modifying one or more Flowering Locus T (FT) genes in a plant and methods of using the compositions for producing plants having suppressed or delayed flowering time. In some embodiments, the plant is a Saccharum plant. The approach used herein involves methods for decreasing expression of one or more Flowering Locus T (FT) genes by RNAi inhibition or CRISPR/Cas9 targeted mutagenesis that results in suppressed or delayed flowering in Saccharum plants, such as sugarcane or energycane. Target genes were isolated from the sugarcane and energycane cultivar and a conserved sequence between these cultivars and closely related species was chosen to design intron-hairpin RNA constructs for RNAi suppression or sgRNA expression constructs for targeted mutagenesis. Recombinant DNA vectors were introduced into sugarcane by biolistic gene transfer and transgenic plants were regenerated and selected, vegetatively propagated for replicated field testing. These genetically modified plants produce significantly elevated biomass and recoverable sugar yield under replicated field conditions relative to a similar plant in which the FT4, FT8, and/or FT10 gene have not been disrupted, when grown under the same conditions.