Novel Epigenetic Techniques Provided by the CRISPR/Cas9 System

Xie, Nina; Zhou, Yifei; Sun, Qiying; Tang, Beisha

doi:10.1155/2018/7834175

Cited by 61 publications

(43 citation statements)

References 95 publications

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“…The key instruments for epigenome editing are the so-called EpiEditors, artificial proteins constructed from two functional units, one of which is responsible for the delivery of the construct to the targeted genomic region and the other for the local modification of the chromatin state to establish the desired properties. Currently, EpiEditors are mainly created using the catalytically inactive Cas9 protein (dCas9) fused to chromatin-modifying enzymes [5]. To produce a functional DNA binding unit, dCas9 needs to form a complex with a single guide RNA (sgRNA), which is designed to contain a sequence complementary to the targeted genomic locus next to a protospacer adjacent motif site ( Figure 1A) [6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Engineering of Effector Domains for Targeted DNA Methylation with Reduced Off-Target Effects

Hofacker

Broche

Laistner

et al. 2020

IJMS

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Epigenome editing is a promising technology, potentially allowing the stable reprogramming of gene expression profiles without alteration of the DNA sequence. Targeted DNA methylation has been successfully documented by many groups for silencing selected genes, but recent publications have raised concerns regarding its specificity. In the current work, we developed new EpiEditors for programmable DNA methylation in cells with a high efficiency and improved specificity. First, we demonstrated that the catalytically deactivated Cas9 protein (dCas9)-SunTag scaffold, which has been used earlier for signal amplification, can be combined with the DNMT3A-DNMT3L single-chain effector domain, allowing for a strong methylation at the target genomic locus. We demonstrated that off-target activity of this system is mainly due to untargeted freely diffusing DNMT3A-DNMT3L subunits. Therefore, we generated several DNMT3A-DNMT3L variants containing mutations in the DNMT3A part, which reduced their endogenous DNA binding. We analyzed the genome-wide DNA methylation of selected variants and confirmed a striking reduction of untargeted methylation, most pronounced for the R887E mutant. For all potential applications of targeted DNA methylation, the efficiency and specificity of the treatment are the key factors. By developing highly active targeted methylation systems with strongly improved specificity, our work contributes to future applications of this approach.

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Section: Introductionmentioning

confidence: 99%

Engineering of Effector Domains for Targeted DNA Methylation with Reduced Off-Target Effects

Hofacker

Broche

Laistner

et al. 2020

IJMS

View full text Add to dashboard Cite

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“…Recent studies have shown the correlation between epigenetics and development and progression of cervical cancer. Epigenetic modifications, such as DNA methylation, hydroxymethylation, demethylation, chromatin remodeling, histone modification, regulation by non-coding RNAs, and gene imprinting, are inheritable and affect genetic information without interfering with mitosis or meiosis (Xie et al, 2018). Among the epigenetic modifications, the role of candidate gene DNA methylation in cervical cancer has been studied the most.…”

Section: Introductionmentioning

confidence: 99%

DNA Methylation and Hydroxymethylation in Cervical Cancer: Diagnosis, Prognosis and Treatment

Zhu

Tian

et al. 2020

Front. Genet.

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Recent discoveries have led to the development of novel ideas and techniques that have helped elucidate the correlation between epigenetics and tumor biology. Nowadays, the field of tumor genetics has evolved to include a new type of regulation by epigenetics. An increasing number of studies have demonstrated the importance of DNA methylation and hydroxymethylation in specific genes in the progression of cervical cancer. Determining the methylation and hydroxymethylation profiles of these genes will help in the early prevention and diagnosis, monitoring recurrence, prognosis, and treatment of patients with cervical cancer. In this review, we focus on the significance of aberrant DNA methylation and hydroxymethylation in cervical cancer and the use of these epigenetic signatures in clinical settings.

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“…A RASSF member, which contains a C-terminal RA domain and is bound to Ras, is the novel Ras effector NORE1 (RASSF5) [58]. It is interesting to note that mutations of Ras oncogenes do not play a major role in the initiation and progression of renal carcinomas [59]. This suggests that the activation of KRAS signaling may be accomplished by the inactivation of RASSF10.…”

Section: Discussionmentioning

confidence: 99%

RASSF10 is frequently epigenetically inactivated in kidney cancer and its knockout promotes neoplasia in cancer prone mice

et al. 2020

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Kidney cancer incidences are rising globally, thereby fueling the demand for targeted therapies and precision medicine. In our previous work, we have identified and characterized the Ras-Association Domain Family encoding ten members that are often aberrantly expressed in human cancers. In this study, we created and analyzed the Rassf10 knockout mice. Here we show that Rassf10 haploinsufficiency promotes neoplasia formation in two established mouse cancer models (Rassf1A −/− and p53 −/−). Haploinsufficient Rassf10 knockout mice were significantly prone to various diseases including lymphoma (Rassf1A −/− background) and thymoma (p53 −/− background). Especially Rassf10 −/− and p53-deficient mice exhibited threefold increased rates of kidney cysts compared with p53 −/− controls. Moreover, we observed that in human kidney cancer, RASSF10 is frequently epigenetically inactivated by its CpG island promoter hypermethylation. Primary tumors of renal clear cell and papillary cell carcinoma confirmed that RASSF10 methylation is associated with decreased expression in comparison to normal kidney tissue. In independent data sets, we could validate that RASSF10 inactivation clinically correlated with decreased survival and with progressed disease state of kidney cancer patients and polycystic kidney size. Functionally, we revealed that the loss of Rassf10 was significantly associated with upregulation of KRAS signaling and MYC expression. In summary, we could show that Rassf10 functions as a haploinsufficient tumor suppressor. In combination with other markers, RASSF10 silencing can serve as diagnostic and prognostic cancer biomarker in kidney diseases.

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Novel Epigenetic Techniques Provided by the CRISPR/Cas9 System

Cited by 61 publications

References 95 publications

Engineering of Effector Domains for Targeted DNA Methylation with Reduced Off-Target Effects

Engineering of Effector Domains for Targeted DNA Methylation with Reduced Off-Target Effects

DNA Methylation and Hydroxymethylation in Cervical Cancer: Diagnosis, Prognosis and Treatment

RASSF10 is frequently epigenetically inactivated in kidney cancer and its knockout promotes neoplasia in cancer prone mice

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