ArticleNucleic acids research2021
Optogenetic control of Neisseria meningitidis Cas9 genome editing using an engineered, light-switchable anti-CRISPR protein.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 43 citations in OpenAlex.
- Optimized optogenetic anti-CRISPR for endogenous gene regulation in Drosophila.Nucleic acids research · 2026Article
- Modular engineering of thermoresponsive allosteric proteins.Nature chemical biology · 2026Article
- Interrogating Physiological Functions with Light and Chemicals.Annual review of physiology · 2026Review
- Article
- Engineering the Link: From Genome Interaction Maps to Functional Insight.Advanced biology · 2025Review
- Rational engineering of allosteric protein switches by in silico prediction of domain insertion sites.Nature methods · 2025Article
- Engineering stimuli-responsive CRISPR-Cas systems for versatile biosensing.Analytical and bioanalytical chemistry · 2025Review
- A modular toolbox for the optogenetic deactivation of transcription.Nucleic acids research · 2025Article
- Protein design accelerates the development and application of optogenetic tools.Computational and structural biotechnology journal · 2025Review
- A deep mutational scanning platform to characterize the fitness landscape of anti-CRISPR proteins.Nucleic acids research · 2024Article
- Rapid characterization of anti-CRISPR proteins and optogenetically engineered variants using a versatile plasmid interference system.Nucleic acids research · 2023Article
- Dissecting the Determinants of Domain Insertion Tolerance and Allostery in Proteins.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- Inhibitory mechanism of CRISPR-Cas9 by AcrIIC4.Nucleic acids research · 2023Article
- Recent advances in therapeutic CRISPR-Cas9 genome editing: mechanisms and applications.Molecular biomedicine · 2023Review
- Optimization of CRISPR-Cas system for clinical cancer therapy.Bioengineering & translational medicine · 2023Review
- LOV2-based photoactivatable CaMKII and its application to single synapses: Local Optogenetics.Biophysics and physicobiology · 2023Article
- Beyond the promise: evaluating and mitigating off-target effects in CRISPR gene editing for safer therapeutics.Frontiers in bioengineering and biotechnology · 2023Review
- Photocontrolled crRNA activation enables robust CRISPR-Cas12a diagnostics.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Design and engineering of light-sensitive protein switches.Current opinion in structural biology · 2022Review
- Controlling CRISPR with small molecule regulation for somatic cell genome editing.Molecular therapy : the journal of the American Society of Gene Therapy · 2022Review
Corrections and comments
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Authors and funding
10 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Optogenetic control of CRISPR-Cas9 systems has significantly improved our ability to perform genome perturbations in living cells with high precision in time and space. As new Cas orthologues with advantageous properties are rapidly being discovered and engineered, the need for straightforward strategies to control their activity via exogenous stimuli persists. The Cas9 from Neisseria meningitidis (Nme) is a particularly small and target-specific Cas9 orthologue, and thus of high interest for in vivo genome editing applications. Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein. Building on our previous Acr engineering work, we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa. Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation. Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface. Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.