The Krüppel-associated Box Repressor Domain Can Induce Reversible Heterochromatization of a Mouse Locus in Vivo

Groner, Anna C.; Tschopp, Patrick; Challet, Ludivine; Dietrich, Jens Erik; Verp, Sonia; Offner, Sandra; Barde, Isabelle; Rodríguez, Iván; Hiiragi, Takashi; Trono, Didier

doi:10.1074/jbc.m112.350884

Cited by 16 publications

(26 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This irreversible silencing was due to KRAB-induced de novo DNA methylation of the promoter ( 13 ). However, a recent study suggested that tethering of KRAB to an endogenous gene body does not contribute to irreversible gene silencing, even under the same circumstances as previously described, as KRAB binding to gene bodies did not induce stable DNA promoter methylation ( 14 ). It is worthy to note that in these experiments, tetO-controlled GFP was inserted into the genome using a lentivirus vector, and in some cases, KAP1 has silenced retroviruses in embryonic stem (ES) cells and early embryos ( 15 – 18 ).…”

Section: Introductionmentioning

confidence: 76%

The Krüppel-associated box repressor domain induces reversible and irreversible regulation of endogenous mouse genes by mediating different chromatin states

Yue

Zhao

Mao

et al. 2015

Nucleic Acids Research

View full text Add to dashboard Cite

The Krüppel-associated box (KRAB) domain is a transcription repression module from the largest family of transcriptional regulators encoded by higher vertebrates. We developed a drug-controllable regulation system based on an artificial KRAB-containing repressor (tTS) that targets the endogenous Hprt gene to explore the regulatory mechanism and molecular basis of KRAB-containing regulators within the context of an endogenous gene in vivo. We show that KRAB can mediate irreversible and reversible regulation of endogenous genes in mouse that is dependent on embryonic developmental stage. KRAB-induced stable DNA methylation within the KRAB binding region during the early embryonic stage, resulting in irreversible gene repression. In later stages, KRAB mainly induced de-acetylation and methylation of histone, resulting in reversible gene repression. Thus, we have characterized the KRAB-mediated regulation system within the context of an endogenous gene and multiple spatiotemporal ranges, thereby providing a basis for identifying the function of KRAB-containing regulators and aiding development of novel KRAB-based gene regulation tools in vivo.

show abstract

Section: Introductionmentioning

confidence: 76%

The Krüppel-associated box repressor domain induces reversible and irreversible regulation of endogenous mouse genes by mediating different chromatin states

Yue

Zhao

Mao

et al. 2015

Nucleic Acids Research

View full text Add to dashboard Cite

show abstract

“…With respect to repression, it has been shown in embryonic stem cells that KRAB (as part of SKD) can indirectly induce de novo DNA methylation via recruiting its cofactor, KAP1 (43). In addition, KRAB/KAP1 can spread H3K9me3-containing heterochromatin in adult and embryonic stem cells (44). Despite these reported functions of KRAB in epigenetic regulation, KRAB-induced repression is generally considered to be transient.…”

Section: Discussionmentioning

confidence: 99%

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

Falahi

Huisman

Kazemier

et al. 2013

Molecular Cancer Research

View full text Add to dashboard Cite

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.

show abstract

“…Synthetic TAL DBDs (T-DBDs) can be appended at either their C-or N-termini with effector domains conferring function in mammalian cells -for example the KRAB repressor domain 15,23 . In both native and artificial TFs, KRAB domains recruit the KAP1 co-repressor and, in turn, endogenous enzymatic complexes that methylate histones and DNA and trigger focal heterochromatin formation [23][24][25][26][27][28] .…”

Section: Resultsmentioning

confidence: 99%

Quantitative dialing of gene expression via precision targeting of KRAB repressor

Wilken

Ciarlo

Pearl

et al. 2020

Preprint

View full text Add to dashboard Cite

Non-invasive epigenome editing is a promising strategy for engineering gene expression programs, yet potency, specificity, and persistence remain challenging. Here we show that effective epigenome editing is gated at single-base precision via 'keyhole' sites in endogenous regulatory DNA. Synthetic repressors targeting promoter keyholes can ablate gene expression in up to 99% of primary cells with single-gene specificity and can seamlessly repress multiple genes in combination. Transient exposure of primary T cells to keyhole repressors confers mitotically heritable silencing that persists to the limit of primary cultures in vitro and for at least 4 weeks in vivo, enabling manufacturing of cell products with enhanced therapeutic efficacy. DNA recognition and effector domains can be encoded as separate proteins that reassemble at keyhole sites and function with the same efficiency as single chain effectors, enabling gated control and rapid screening for novel functional domains that modulate endogenous gene expression patterns. Our results provide a powerful and exponentially flexible system for programming gene expression and therapeutic cell products.Here we report that the potency and specificity of epigenetic transcriptional repression are linked through promoter keyhole sites, the targeting of which triggers nearcomplete repression with single-gene accuracy. Potent repression, in turn, enables durable programming via reliable induction of mitotically heritable expression programs, offering new potential for engineered cell therapies. 7

show abstract

The Krüppel-associated Box Repressor Domain Can Induce Reversible Heterochromatization of a Mouse Locus in Vivo

Cited by 16 publications

References 40 publications

The Krüppel-associated box repressor domain induces reversible and irreversible regulation of endogenous mouse genes by mediating different chromatin states

The Krüppel-associated box repressor domain induces reversible and irreversible regulation of endogenous mouse genes by mediating different chromatin states

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

Quantitative dialing of gene expression via precision targeting of KRAB repressor

Contact Info

Product

Resources

About