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Abstract
The novel single-guide RNA architecture achieved up to 215-fold induction of three endogenous rice genes in protoplast assays.
- CRISPR activation (CRISPRa) can effectively upregulate endogenous genes in plants.
- A new sgRNA configuration with two MS2 aptamers and a specific promoter showed significant improvement over previous constructs.
- Scaffold RNAs with improper aptamer positioning or excessive copies were not effective in gene activation.
- Exploratory modeling suggests that aptamer arrangement and protein dimerization may influence activation efficiency.
- The design allows flexible gene regulation in rice protoplasts and is suitable for high-throughput applications.
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