Journal article
Simulations Predict Stronger CRISPRi Transcriptional Repression in Plants for Identical than Heterogeneous gRNA Target Sites
Synthetic biology (Oxford University Press), Vol.10(1), ysae020
04/14/2025
DOI: 10.1093/synbio/ysae020
PMCID: PMC12007490
PMID: 40255684
Abstract
Plant synthetic biologists have been working to adapt the CRISPRa and CRISPRi promoter regulation methods for applications such as improving crops or installing other valuable pathways. With other organisms, strong transcriptional control has typically required multiple gRNA target sites, which poses a critical engineering choice between heterogeneous sites, which allow each gRNA to target existing locations in a promoter, and identical sites, which typically require modification of the promoter. Here, we investigate the consequences of this choice for CRISPRi plant promoter regulation via simulation-based analysis, using model parameters based on single gRNA regulation and constitutive promoters in Nicotiana benthamiana and Arabidopsis thaliana. Using models of 2 to 6 gRNA target sites to compare heterogeneous versus identical sites for tunability, sensitivity to parameter values, and sensitivity to cell-to-cell variation, we find that identical gRNA target sites are predicted to yield far more effective transcriptional repression than heterogeneous sites.
Details
- Title: Subtitle
- Simulations Predict Stronger CRISPRi Transcriptional Repression in Plants for Identical than Heterogeneous gRNA Target Sites
- Creators
- Helen ScottAlessandro OcchialiniScott C LenaghanJacob Beal
- Resource Type
- Journal article
- Publication Details
- Synthetic biology (Oxford University Press), Vol.10(1), ysae020
- DOI
- 10.1093/synbio/ysae020
- PMID
- 40255684
- PMCID
- PMC12007490
- NLM abbreviation
- Synth Biol (Oxf)
- ISSN
- 2397-7000
- eISSN
- 2397-7000
- Publisher
- OXFORD UNIV PRESS; OXFORD
- Grant note
- DARPA: HR0011-18-2-0049
This work was supported by DARPA contract HR0011-18-2-0049. This document does not contain technology or technical data controlled under either U.S. International Traffic in Arms Regulation or US Export Administration Regulations. Views, opinions, and/or findings expressed are those of the author(s) and should not be interpreted as representing the official views or policies of the Department of Defense or the US Government. This study by approved for public release, distribution is unlimited (DISTAR Case 39302).
- Language
- English
- Date published
- 04/14/2025
- Academic Unit
- Electrical and Computer Engineering
- Record Identifier
- 9984811212802771
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