G-quadruplex-based CRISPR photoswitch for spatiotemporal control of genomic modulation
Open accessAbstract
CRISPR (clustered regularly interspaced short palindromic repeats) technology holds tremendous promise for gene regulation and editing. However, precise control of CRISPR editing is essential to overcome its uncontrollable reaction process and excessive activity that leads to off-target editing. To overcome this problem, we engineered a photoswitch on G-quadruplex gRNA (GqRNA) for precisely controlled gene editing and expression by embedding dicationic azobenzene derivatives (AZD++). Our results demonstrated that rational design of the G-quadruplex onto crRNA conferred higher stability and sequence recognition specificity than unmodified single guide (sgRNA). Light-induced isomerization of AZD++ quickly transformed the on state of GqRNA, which facilitated rapid activation of ribonucleoprotein activity for genome editing of on-target sites in cells with excellent editing efficiency. In turn, AZD++–GqRNA promptly refolded to an off state to inhibit genomic cleavage, and limited the generation of off-target effects and by-products. Therefore, the proposed strategy of a photo-reversible modality presents a new opportunity for CRISPR-Cas9 modulation to improve its safety and applicability.
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Controlled CRISPR approaches substantially mitigate off-target risks by enabling more targeted genetic interventions.
“Such controlled approaches substantially mitigate off-target risks by enabling more targeted genetic interventions (Deng et al., 2023).”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Deng et al., 2023
G-quadruplex-based CRISPR photoswitches provide spatiotemporal control over CRISPR activity, allowing researchers to precisely activate and deactivate the CRISPR system.
“These innovative mechanisms provide spatiotemporal control over CRISPR activity, allowing researchers to precisely activate and deactivate the CRISPR system.”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Deng et al., 2023
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Engineering Guide RNAs for CRISPR-Based Biosensors
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- Authors
- Deng, H., Xu, H., Wang, Y., Jia, R., Ma, X., Feng, Y., & Chen, H.
- Affiliations
- Xiamen University (CN)
- State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics & Center for Molecular Imaging and Translational Medicine, School of Public Health, Xiamen University , Xiamen 361102 , China
- Corresponding author
- Chen, Hongmin
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2023-03-13
- Volume
- 51
- Issue
- 8
- Pages
- 4064-4077
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC10164585
- OpenAlex
- W4324021335
- Cited by
- 45
- References
- 60
- Citations by year
- 2026: 19 · 2025: 16 · 2024: 7 · 2023: 3
- FWCI
- 4.01
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Nucleic Acids Research (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- National Natural Science Foundation of China (82172007, 81771977)
- Distinguished Young Scholars of Fujian Province (2021J06007)
- Xiamen Science and Technology Plan Project (3502Z20183017)
- Fundamental Research Funds for the Central Universities of China (20720180054)
- National Facility for Translational Medicine (TMSK-2021-102)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Physical Sciences › Materials Science › Materials Chemistry › Photochromic and Fluorescence Chemistry
- Keywords
- CRISPR/Cas9, G-quadruplex, photoswitches, azobenzene, spatiotemporal control, off-target reduction, single guide RNA, photoisomerization, gene expression regulation
- MeSH terms
- Gene Editing, Gene Expression Regulation, Genome, Genomics, CRISPR-Cas Systems
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