Targeted Methylation and Gene Silencing of VEGF-A in Human Cells by Using a Designed Dnmt3a–Dnmt3L Single-Chain Fusion Protein with Increased DNA Methylation Activity

Siddique, Abu Nasar; Nunna, Suneetha; Rajavelu, Arumugam; Zhang, Yingying; Jurkowska, Renata Z.; Reinhardt, Richard; Rots, Marianne G.; Ragozin, Sergey; Jurkowski, Tomasz P.; Jeltsch, Albert

doi:10.1016/j.jmb.2012.11.038

Cited by 143 publications

(120 citation statements)

References 64 publications

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“…Further cellular studies are now required to investigate the physiological relevance of DNMT-3A and -3B as players in active DNA demethylation. Epigenetic editing tools [51][52][53][54][55] offer exciting possibilities to address differential effects of epigenetic enzymes in various chromatin contexts in living cells. 60 …”

Section: Resultsmentioning

confidence: 99%

“…To identify such situations in physiological situations, the targeting of DNMTs to pre-determined genomic loci by epigenetic editing, as described by us and others previously [51][52][53][54][55] provides an interesting tool to test and to eventually exploit DNA demethylation activities of DNMTs in vivo.…”

Section: Dnmt-3a and -3b As Demethylating Enzymes: Implicationsmentioning

confidence: 99%

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Local chromatin microenvironment determines DNMT activity: from DNA methyltransferase to DNA demethylase or DNA dehydroxymethylase

et al. 2015

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Section: Resultsmentioning

confidence: 99%

Section: Dnmt-3a and -3b As Demethylating Enzymes: Implicationsmentioning

confidence: 99%

Local chromatin microenvironment determines DNMT activity: from DNA methyltransferase to DNA demethylase or DNA dehydroxymethylase

et al. 2015

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“…Modulation of expression of endogenous genes by epigenetic editing would open up exciting venues. Targeted DNA methylation to repress gene expression was recently reported for three endogenous genes (28,29). Inducing repressive histone modifications has been reported for one gene; upon targeting of catalytic domains of histone methyltransferase enzymes (G9a, SUV39-H1) to the VEGF-A promoter, repressive histone marks were induced and result in downregulation of gene expression (30).…”

Section: Introductionmentioning

confidence: 99%

Towards Sustained Silencing of HER2/neu in Cancer By Epigenetic Editing

Falahi

Huisman

Kazemier

et al. 2013

Molecular Cancer Research

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The human epidermal growth factor receptor-2 (HER2/neu/ERBB2) is overexpressed in several cancer types. Although therapies targeting the HER2/neu protein result in inhibition of cell proliferation, the anticancer effect might be further optimized by limiting HER2/neu expression at the DNA level. Towards this aim, epigenetic editing was performed to suppress HER2/neu expression by inducing epigenetic silencing marks on the HER2/neu promoter.HER2/neu expression and HER2/neu promoter epigenetic modification status were determined in a panel of ovarian and breast cancer cell lines. HER2/neu-overexpressing cancer cells were transduced to express a zinc finger protein (ZFP), targeting the HER2/neu gene, fused to histone methyltransferases (G9a, SUV39-H1)/super KRAB domain (SKD). Epigenetic assessment of the HER2/neu promoter showed that HER2/neu-ZFP fused to G9a efficiently induced the intended silencing histone methylation mark (H3K9me2). Importantly, H3K9me2 induction was associated with a dramatic downregulation of HER2/neu expression in HER2/neu-overexpressing cells. Downregulation by SKD, traditionally considered transient in nature, was associated with removal of the histone acetylation mark (H3ac). The downregulation of HER2/neu by induced H3K9 methylation and/or reduced H3 acetylation was sufficient to effectively inhibit cellular metabolic activity and clonogenicity. Furthermore, genome-wide analysis indicated preferential binding of the ZFP to its target sequence. These results not only show that H3K9 methylation can be induced but also that this epigenetic mark was instructive in promoting downregulation of HER2/neu expression.

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“…Among the first fully synthetic transcriptional effector proteins to be generated (Beerli et al 1998) were those based on the fusion of engineered zinc-finger proteins with either the Herpes simplex-derived transactivation domain (Sadowski et al 1988) or the Krüp-pel-associated box (KRAB) repression protein (Margolin et al 1994). Over the course of the next 15 years, zinc-finger-based transcriptional modulators were expanded and featured several other types of effector domains (Beerli and Barbas 2002), including, for example, the Dnmt3a methyltransferase domain (Rivenbark et al 2012;Siddique et al 2013) and the ten-eleven translocation methylcytosine dioxygenase 1 (TET1) , which can modulate transcription via targeted methylation or demethylation, respectively. As a natural extension of zinc-finger transcription factors, and further drawing on the parallels with zinc-finger proteins, TALE transcription factors have also emerged as an especially effective platform for achieving targeted transcriptional modulation Zhang et al 2011).…”

Section: Targeted Transcription Factors Tools For Modulating Gene Expmentioning

confidence: 99%

Genome-Editing Technologies: Principles and Applications

Gaj

Sirk

Shui

et al. 2016

Cold Spring Harb Perspect Biol

270

142

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Targeted nucleases have provided researchers with the ability to manipulate virtually any genomic sequence, enabling the facile creation of isogenic cell lines and animal models for the study of human disease, and promoting exciting new possibilities for human gene therapy. Here we review three foundational technologies-clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9), transcription activator-like effector nucleases (TALENs), and zinc-finger nucleases (ZFNs). We discuss the engineering advances that facilitated their development and highlight several achievements in genome engineering that were made possible by these tools. We also consider artificial transcription factors, illustrating how this technology can complement targeted nucleases for synthetic biology and gene therapy.

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Targeted Methylation and Gene Silencing of VEGF-A in Human Cells by Using a Designed Dnmt3a–Dnmt3L Single-Chain Fusion Protein with Increased DNA Methylation Activity

Cited by 143 publications

References 64 publications

Local chromatin microenvironment determines DNMT activity: from DNA methyltransferase to DNA demethylase or DNA dehydroxymethylase

Local chromatin microenvironment determines DNMT activity: from DNA methyltransferase to DNA demethylase or DNA dehydroxymethylase

Towards Sustained Silencing of HER2/neu in Cancer By Epigenetic Editing

Genome-Editing Technologies: Principles and Applications

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