SB-Actin Terminator Sequence for Plant Gene Expression
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Solution Overview
Problem
Current plant genetic engineering techniques face challenges in modulating gene expression in transgenic plants due to leaky transcription termination and bidirectional, convergent transcription, which can lead to undesirable effects such as decreased transgene expression and antisense RNA production, highlighting the need for efficient and novel transcriptional terminators.
Innovation Solution
The use of novel terminator sequences, such as the SB-ACTIN terminator, which are designed to function effectively in plant cells to regulate gene expression by preventing read-through transcription and ensuring optimal expression of heterologous polynucleotides, are integrated into recombinant DNA constructs and expressed in plants to control gene expression accurately.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional terminator sequences are used in transgenic plants, then gene expression can be regulated, but leaky transcription termination occurs causing decreased transgene expression and antisense RNA production
Solution Approach 1:
The patent applies parameter changes by modifying the terminator sequence parameters - specifically using the SB-actin terminator sequence from sorghum instead of conventional terminators. This sequence parameter change results in improved transcription termination efficiency and eliminates the harmful leaky termination effects that cause antisense RNA production. The SB-actin terminator demonstrates superior performance in preventing read-through transcription compared to conventional terminators.
Solution Approach 2:
The patent employs the copying principle by utilizing the naturally occurring SB-actin terminator sequence from sorghum genome and replicating it for use in transgenic plant systems. This copied natural terminator sequence proves more effective than artificially designed or conventional terminators, showing that copying proven natural regulatory elements can resolve transcription termination issues.
2Adaptability or versatility
If multiple transgenes are introduced into plants for gene stacking, then complex phenotypes can be achieved, but repetitive sequences cause negative effects on transgene expression and stability
Solution Approach 1:
The patent applies local quality by using diverse regulatory elements with distinct sequence characteristics for different transgenes. Specifically, the SB-actin terminator provides a unique regulatory sequence composition that differs from other terminators, allowing each transgene to have optimized local regulatory properties. This prevents the negative effects of repetitive sequences while maintaining multi-trait expression capability.
Solution Approach 2:
The patent employs composite materials principle by constructing transgenic plants with a mosaic of diverse regulatory elements from different sources. The SB-actin terminator, being a composite regulatory sequence with unique structural and functional properties, is combined with other diverse regulatory elements to create a stable multi-trait expression system that avoids the instability caused by repetitive sequences.
3Productivity
If bidirectional and convergent transcription occurs in transgenic plants, then multiple genes can be expressed, but transcription read-through decreases expression levels and generates unwanted RNA
Solution Approach 1:
The patent applies preliminary anti-action by using the SB-actin terminator sequence that is specifically designed to prevent read-through transcription before it can occur. This terminator sequence acts in advance to block improper transcription elongation, thereby preventing the formation of harmful read-through transcripts and maintaining high expression levels of intended transgenes.
Data Source
AI summary
The present invention discloses polynucleotide sequences that can be used to regulate gene expression in plants. Terminator sequences from Sorghum bicolor that are functional in plants are disclosed. Nucleic acid molecules, recombinant expression constructs, plants and seed comprising these terminator sequences are further disclosed.