ArticleNucleic acids research2019
Synthetic switch to minimize CRISPR off-target effects by self-restricting Cas9 transcription and translation.
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 38 papers.
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Who cites it
38 citing papers in PubMed.
- Improvement of chicken genome editing efficiency in vitro using ribonucleoprotein-mediated CRISPR/Cas9 delivery.Journal of animal science and biotechnology · 2026Article
- VaxLab: integrated platform for rapid multistrategy mRNA vaccine design.Experimental & molecular medicine · 2026Article
- CRISPR-associated transposon for programmable viral vector engineering and prime editing.Nucleic acids research · 2026Article
- Advancements in Osteosarcoma Therapy: Overcoming Chemotherapy Resistance and Exploring Novel Pharmacological Strategies.Pharmaceuticals (Basel, Switzerland) · 2025Review
- Tumor cell membrane-based vaccines: A potential boost for cancer immunotherapy.Exploration (Beijing, China) · 2024Review
- Intelligent guide RNA: dual toehold switches for modulating luciferase in the presence of trigger RNA.Communications biology · 2024Article
- Expression of microbial lipase in filamentous fungus3 Biotech · 2024Review
- Recent Research Trends in Stem Cells Using CRISPR/Cas-Based Genome Editing Methods.International journal of stem cells · 2024Review
- Comprehensive review of CRISPR-based gene editing: mechanisms, challenges, and applications in cancer therapy.Molecular cancer · 2024Review
- Enhancement of specialized metabolites using CRISPR/Cas gene editing technology in medicinal plants.Frontiers in plant science · 2024Review
- Trends and challenges in organoid modeling and expansion with pluripotent stem cells and somatic tissue.PeerJ · 2024Review
- CRISPR/Cas9: a powerful tool in colorectal cancer research.Journal of experimental & clinical cancer research : CR · 2023Review
- An overview of genome engineering in plants, including its scope, technologies, progress and grand challenges.Functional & integrative genomics · 2023Review
- Genome Editing and Pathological Cardiac Hypertrophy.Advances in experimental medicine and biology · 2023Article
- CRISPR/dCas9 for hepatic fibrosis therapy: implications and challenges.Molecular biology reports · 2022Review
- Therapeutic Applications of the CRISPR-Cas System.Bioengineering (Basel, Switzerland) · 2022Review
- CRISPR/Cas9-Based Genome Editing and Its Application inJournal of fungi (Basel, Switzerland) · 2022Review
- Directed evolution of orthogonal RNA-RBP pairs through library-vs-library in vitro selection.Nucleic acids research · 2022Article
- Effect of CRISPR/Cas9-Edited PD-1/PD-L1 on Tumor Immunity and Immunotherapy.Frontiers in immunology · 2022Review
- Protein-Based Systems for Translational Regulation of Synthetic mRNAs in Mammalian Cells.Life (Basel, Switzerland) · 2021Review
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR/Cas9 is a powerful genome editing system but uncontrolled Cas9 nuclease expression triggers off-target effects and even in vivo immune responses. Inspired by synthetic biology, here we built a synthetic switch that self-regulates Cas9 expression not only in the transcription step by guide RNA-aided self-cleavage of cas9 gene, but also in the translation step by L7Ae:K-turn repression system. We showed that the synthetic switch enabled simultaneous transcriptional and translational repression, hence stringently attenuating the Cas9 expression. The restricted Cas9 expression induced high efficiency on-target indel mutation while minimizing the off-target effects. Furthermore, we unveiled the correlation between Cas9 expression kinetics and on-target/off-target mutagenesis. The synthetic switch conferred detectable Cas9 expression and concomitant high frequency on-target mutagenesis at as early as 6 h, and restricted the Cas9 expression and off-target effects to minimal levels through 72 h. The synthetic switch is compact enough to be incorporated into viral vectors for self-regulation of Cas9 expression, thereby providing a novel 'hit and run' strategy for in vivo genome editing.
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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.