ArticleNucleic acids research2023
Phase-separated DropCRISPRa platform for efficient gene activation in mammalian cells and mice.
Article in Nucleic acids research, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 26 citations in OpenAlex.
- NS1-Induced PARP1 Phase Transitions Prolongs Host Transcriptional Pausing to Promote Influenza A Virus Replication.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Advances in engineering and applications of synthetic phase-separated membraneless organelles in biotechnology.Synthetic and systems biotechnology · 2026Review
- Programmable nanobody circuits for cell selection.bioRxiv : the preprint server for biology · 2026Article
- Programmable transcriptional condensates for enhanced CRISPR-based gene regulation.Theranostics · 2026Article
- Engineered membraneless organelles inSynthetic and systems biotechnology · 2025Article
- Osmotic pressure regulates DNA labelling and transcription with dCas9-SunTag system in live cells.Nature communications · 2025Article
- Liquid-Liquid Phase Separation: Mechanisms, Roles, and Implications in Cellular Function and Disease.FASEB bioAdvances · 2025Review
- hpCasMINI: An engineered hypercompact CRISPR-Cas12f system with boosted gene editing activity.Nature communications · 2025Article
- Engineered circular guide RNAs enhance miniature CRISPR/Cas12f-based gene activation and adenine base editing.Nature communications · 2025Article
- Dynamic properties of transcriptional condensates modulate CRISPRa-mediated gene activation.Nature communications · 2025Article
- Biomolecular condensation of human IDRs initiates endogenous transcription via intrachromosomal looping or high-density promoter localization.Nucleic acids research · 2025Article
- Liquid-liquid phase separation: an emerging perspective on the tumorigenesis, progression, and treatment of tumors.Frontiers in immunology · 2025Review
- A sensitive red/far-red photoswitch for controllable gene therapy in mouse models of metabolic diseases.Nature communications · 2024Article
- Biomolecular condensates and disease pathogenesis.Science China. Life sciences · 2024Review
- Specific multivalent molecules boost CRISPR-mediated transcriptional activation.Nature communications · 2024Article
- Expanding RNA editing toolkit using an IDR-based strategy.Molecular therapy. Nucleic acids · 2024Article
- Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor.Nucleic acids research · 2024Article
- Technologies for studying phase-separated biomolecular condensates.Advanced biotechnology · 2024Review
Corrections and comments
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Authors and funding
12 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Liquid-liquid phase separation (LLPS) plays a critical role in regulating gene transcription via the formation of transcriptional condensates. However, LLPS has not been reported to be engineered as a tool to activate endogenous gene expression in mammalian cells or in vivo. Here, we developed a droplet-forming CRISPR (clustered regularly interspaced short palindromic repeats) gene activation system (DropCRISPRa) to activate transcription with high efficiency via combining the CRISPR-SunTag system with FETIDR-AD fusion proteins, which contain an N-terminal intrinsically disordered region (IDR) of a FET protein (FUS or TAF15) and a transcription activation domain (AD, VP64/P65/VPR). In this system, the FETIDR-AD fusion protein formed phase separation condensates at the target sites, which could recruit endogenous BRD4 and RNA polymerase II with an S2 phosphorylated C-terminal domain (CTD) to enhance transcription elongation. IDR-FUS9Y>S and IDR-FUSG156E, two mutants with deficient and aberrant phase separation respectively, confirmed that appropriate phase separation was required for efficient gene activation. Further, the DropCRISPRa system was compatible with a broad set of CRISPR-associated (Cas) proteins and ADs, including dLbCas12a, dAsCas12a, dSpCas9 and the miniature dUnCas12f1, and VP64, P65 and VPR. Finally, the DropCRISPRa system could activate target genes in mice. Therefore, this study provides a robust tool to activate gene expression for foundational research and potential therapeutics.
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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.