19 publications found.

Analysis of estrogen-regulated enhancer RNAs identifies a functionalmotif required for enhancer assembly and gene expression.
Published: June, 2022

Abstract: To better understand the functions of non-coding enhancer RNAs (eRNAs), we annotated the estrogen-regulated eRNA transcriptome in estrogen receptor α (ERα)-positive breast cancer cells using PRO-cap and RNA sequencing. We then cloned a...
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Antisense non-coding transcription represses the PHO5 model genevia remodelling of promoter chromatin structure
Published: February, 2022

Abstract: Pervasive transcription of eukaryotic genomes generates non-coding transcripts with regulatory potential. We examined the effects of non-coding antisense transcription on the regulation of expression of the yeast PHO5 gene, a paradigmatic case for...
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A predominant enhancer co-amplified with the oncogene is necessary andsufficient for its expression in squamous cancer
Published: December, 2021

Abstract: Amplification and overexpression of the SOX2 oncogene represent a hallmark of squamous cancers originating from diverse tissue types. Here, we find that squamous cancers selectively amplify a 3’ noncoding region together with SOX2, which har...
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Establishment of a second generation homozygous CRISPRa human inducedpluripotent stem cell (hiPSC) line for enhanced levels of endogenous geneactivation.
Published: October, 2021

Abstract: CRISPR/Cas9 technology based on nuclease inactive dCas9 and fused to the heterotrimeric VPR transcriptional activator is a powerful tool to enhance endogenous transcription by targeting defined genomic loci. We generated homozygous human induced p...
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MICAL1 regulates actin cytoskeleton organization, directional cellmigration and the growth of human breast cancer cells as orthotopicxenograft tumours.
Published: October, 2021

Abstract: The Molecule Interacting with CasL 1 (MICAL1) monooxygenase has emerged as an important regulator of cytoskeleton organization via actin oxidation. Although filamentous actin (F-actin) increases MICAL1 monooxygenase activity, hydrogen peroxide (HO...
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Establishment of two homozygous CRISPR interference (CRISPRi)knock-in human induced pluripotent stem cell (hiPSC) lines for titratableendogenous gene repression.
Published: August, 2021

Abstract: Using nuclease-deficient dead (d)Cas9 without enzymatic activity fused to transcriptional inhibitors (CRISPRi) allows for transcriptional interference and results in a powerful tool for the elucidation of developmental, homeostatic and disease mec...
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Transgenic mice for in vivo epigenome editing with CRISPR-based systems
Published: March, 2021

Abstract: The discovery, characterization, and adaptation of the RNA-guided clustered regularly interspersed short palindromic repeat (CRISPR)-Cas9 system has greatly increased the ease with which genome and epigenome editing can be performed. Fusion of chr...
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TGFβ promotes widespread enhancer chromatin opening and operates ongenomic regulatory domains.
Published: December, 2020

Abstract: The Transforming Growth Factor-β (TGFβ) signaling pathway controls transcription by regulating enhancer activity. How TGFβ-regulated enhancers are selected and what chromatin changes are associated with TGFβ-dependent enhancers...
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A gene therapy for inherited blindness using dCas9-VPR–mediatedtranscriptional activation
Published: August, 2020

Abstract: Catalytically inactive dCas9 fused to transcriptional activators (dCas9-VPR) enables activation of silent genes. Many disease genes have counterparts, which serve similar functions but are expressed in distinct cell types. One attractive option to...
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CRISPR-based gene knockout screens reveal deubiquitinases involved in HIV-1 latency in two Jurkat cell models.
Published: March, 2020

Abstract: The major barrier to a HIV-1 cure is the persistence of latent genomes despite treatment with antiretrovirals. To investigate host factors which promote HIV-1 latency, we conducted a genome-wide functional knockout screen using CRISPR-Cas9 in a HI...
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DNA Methylation Editing by CRISPR-guided Excision of 5-Methylcytosine.
Published: February, 2020

Abstract: Tools for actively targeted DNA demethylation are required to increase our knowledge about regulation and specific functions of this important epigenetic modification. DNA demethylation in mammals involves TET-mediated oxidation of 5-methylcytosin...
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DAPK1 loss triggers tumor invasion in colorectal tumor cells.
Published: November, 2019

Abstract: Colorectal cancer (CRC) is one of the leading cancer-related causes of death worldwide. Despite the improvement of surgical and chemotherapeutic treatments, as of yet, the disease has not been overcome due to metastasis to distant organs. Hence, i...
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Guidelines for optimized gene knockout using CRISPR/Cas9
Published: May, 2019

Abstract: CRISPR/Cas9 technology has evolved as the most powerful approach to generate genetic models both for fundamental and preclinical research. Despite its apparent simplicity, the outcome of a genome-editing experiment can be substantially impacted by...
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Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy.
Published: March, 2019

Abstract: Duchenne muscular dystrophy (DMD) is a monogenic disorder and a candidate for therapeutic genome editing. There have been several recent reports of genome editing in preclinical models of Duchenne muscular dystrophy, however, the long-term persist...
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One-step generation of modular CAR-T cells with AAV-Cpf1.
Published: March, 2019

Abstract: Immune-cell engineering opens new capabilities for fundamental immunology research and immunotherapy. We developed a system for efficient generation of chimeric antigen receptor (CAR)-engineered T cells (CAR-T cells) with considerably enhanced fea...
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CRISPR-mediated activation of a promoter or enhancer rescues obesity caused by haploinsufficiency.
Published: January, 2019

Abstract: A wide range of human diseases result from haploinsufficiency, where the function of one of the two gene copies is lost. Here, we targeted the remaining functional copy of a haploinsufficient gene using CRISPR-mediated activation (CRISPRa) in and ...
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zHSF1 modulates zper2 expression in zebrafish embryos
Published: March, 2018

Abstract: HSF1 is a transcription factor that plays a key role in circadian resetting by temperature. We have used zebrafish embryos to decipher the roles of zHsf1, heat and light on zper2 transcription in vivo. Our results show that heat shock (HS) stimula...
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RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells
Published: January, 2018

Abstract: Regenerative therapies require availability of an abundant healthy cell source which can be achieved by e±cient cryopreservation techniques. Here, we established a novel approach for improved cell cryopreservation using an mRNA-based dCas9-...
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A self-inactivating system for AAV-mediated in vivo base editing

Abstract: DNA base editors have been harnessed as an exciting therapeutic platform for human diseases and are rapidly progressing into human clinical trials. However, persistent expression of base editors delivered via adeno-associated virus (AAV) poses con...
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