ArticleNucleic acids research2019
Achieving tight control of a photoactivatable Cre recombinase gene switch: new design strategies and functional characterization in mammalian cells and rodent.
Article in Nucleic acids research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 40 citations in OpenAlex.
- Programmable promoter editing for precise control of transgene expression.Nature biotechnology · 2026Article
- Bioengineering mini-colons for ex vivo colorectal cancer research.Nature protocols · 2026Review
- A Single-Chain Light-Activatable Transcriptional Reporter for Fluorescently Tagging Mammalian Cells In Vitro.Chembiochem : a European journal of chemical biology · 2026Article
- Article
- Review
- Programmable promoter editing for precise control of transgene expression.bioRxiv : the preprint server for biology · 2024Article
- Article
- Stable Transgenic Mouse Strain with Enhanced Photoactivatable Cre Recombinase for Spatiotemporal Genome Manipulation.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2022Article
- Reprogramming the Cleavage Specificity of Botulinum Neurotoxin Serotype B1.ACS synthetic biology · 2022Article
- Optophysiology: Illuminating cell physiology with optogenetics.Physiological reviews · 2022Review
- Fate-mapping mice: new tools and technology for immune discovery.Trends in immunology · 2022Review
- Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives.Frontiers in bioengineering and biotechnology · 2022Review
- Nuclear Localization Signals for Optimization of Genetically Encoded Tools in Neurons.Frontiers in cell and developmental biology · 2022Article
- Signaling, Deconstructed: Using Optogenetics to Dissect and Direct Information Flow in Biological Systems.Annual review of biomedical engineering · 2021Review
- Optobiochemistry: Genetically Encoded Control of Protein Activity by Light.Annual review of biochemistry · 2021Review
- Dual Systems for Enhancing Control of Protein Activity through Induced Dimerization Approaches.Advanced biology · 2021Review
- Steering Molecular Activity with Optogenetics: Recent Advances and Perspectives.Advanced biology · 2021Review
- A light way for nuclear cell biologists.Journal of biochemistry · 2021Review
- A single-chain and fast-responding light-inducible Cre recombinase as a novel optogenetic switch.eLife · 2021Article
- Light Control of Gene Expression Dynamics.Advances in experimental medicine and biology · 2021Review
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
Abstract
A common mechanism for inducibly controlling protein function relies on reconstitution of split protein fragments using chemical or light-induced dimerization domains. A protein is split into fragments that are inactive on their own, but can be reconstituted after dimerization. As many split proteins retain affinity for their complementary half, maintaining low activity in the absence of an inducer remains a challenge. Here, we systematically explore methods to achieve tight regulation of inducible proteins that are effective despite variation in protein expression level. We characterize a previously developed split Cre recombinase (PA-Cre2.0) that is reconstituted upon light-induced CRY2-CIB1 dimerization, in cultured cells and in vivo in rodent brain. In culture, PA-Cre2.0 shows low background and high induced activity over a wide range of expression levels, while in vivo the system also shows low background and sensitive response to brief light inputs. The consistent activity stems from fragment compartmentalization that shifts localization toward the cytosol. Extending this work, we exploit nuclear compartmentalization to generate light-and-chemical regulated versions of Cre recombinase. This work demonstrates in vivo functionality of PA-Cre2.0, describes new approaches to achieve tight inducible control of Cre DNA recombinase, and provides general guidelines for further engineering and application of split protein fragments.
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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.