Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1
DNA methylation is critically involved in the regulation of chromatin states and cell-type-specific gene expression. The exclusive expression of imprinted genes from either the maternal or the paternal allele is regulated by allele-specific DNA methylation at imprinting control regions (ICRs). Aberr...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-01-01
|
| Series: | Genes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4425/15/1/80 |
| _version_ | 1797343830167519232 |
|---|---|
| author | Claudia Albrecht Nivethika Rajaram Julian Broche Pavel Bashtrykov Albert Jeltsch |
| author_facet | Claudia Albrecht Nivethika Rajaram Julian Broche Pavel Bashtrykov Albert Jeltsch |
| author_sort | Claudia Albrecht |
| collection | DOAJ |
| description | DNA methylation is critically involved in the regulation of chromatin states and cell-type-specific gene expression. The exclusive expression of imprinted genes from either the maternal or the paternal allele is regulated by allele-specific DNA methylation at imprinting control regions (ICRs). Aberrant DNA hyper- or hypomethylation at the ICR1 of the <i>H19/IGF2</i> imprinting locus is characteristic for the imprinting disorders Beckwith–Wiedemann syndrome (BWS) and Silver–Russell syndrome (SRS), respectively. In this paper, we performed epigenome editing to induce targeted DNA demethylation at ICR1 in HEK293 cells using dCas9-SunTag and the catalytic domain of TET1. 5-methylcytosine (5mC) levels at the target locus were reduced up to 90% and, 27 days after transient transfection, >60% demethylation was still observed. Consistent with the stable demethylation of CTCF-binding sites within the ICR1, the occupancy of the DNA methylation-sensitive insulator CTCF protein increased by >2-fold throughout the 27 days. Additionally, the <i>H19</i> expression was increased by 2-fold stably, while <i>IGF2</i> was repressed though only transiently. Our data illustrate the ability of epigenome editing to implement long-term changes in DNA methylation at imprinting control regions after a single transient treatment, potentially paving the way for therapeutic epigenome editing approaches in the treatment of imprinting disorders. |
| first_indexed | 2024-03-08T10:53:27Z |
| format | Article |
| id | doaj.art-792651967a1243c58619181497cd55de |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-08T10:53:27Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-792651967a1243c58619181497cd55de2024-01-26T16:41:55ZengMDPI AGGenes2073-44252024-01-011518010.3390/genes15010080Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1Claudia Albrecht0Nivethika Rajaram1Julian Broche2Pavel Bashtrykov3Albert Jeltsch4Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanyInstitute of Biochemistry and Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanyInstitute of Biochemistry and Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanyInstitute of Biochemistry and Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanyInstitute of Biochemistry and Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, GermanyDNA methylation is critically involved in the regulation of chromatin states and cell-type-specific gene expression. The exclusive expression of imprinted genes from either the maternal or the paternal allele is regulated by allele-specific DNA methylation at imprinting control regions (ICRs). Aberrant DNA hyper- or hypomethylation at the ICR1 of the <i>H19/IGF2</i> imprinting locus is characteristic for the imprinting disorders Beckwith–Wiedemann syndrome (BWS) and Silver–Russell syndrome (SRS), respectively. In this paper, we performed epigenome editing to induce targeted DNA demethylation at ICR1 in HEK293 cells using dCas9-SunTag and the catalytic domain of TET1. 5-methylcytosine (5mC) levels at the target locus were reduced up to 90% and, 27 days after transient transfection, >60% demethylation was still observed. Consistent with the stable demethylation of CTCF-binding sites within the ICR1, the occupancy of the DNA methylation-sensitive insulator CTCF protein increased by >2-fold throughout the 27 days. Additionally, the <i>H19</i> expression was increased by 2-fold stably, while <i>IGF2</i> was repressed though only transiently. Our data illustrate the ability of epigenome editing to implement long-term changes in DNA methylation at imprinting control regions after a single transient treatment, potentially paving the way for therapeutic epigenome editing approaches in the treatment of imprinting disorders.https://www.mdpi.com/2073-4425/15/1/80epigenome editingimprintingDNA demethylationdCas9SunTagTET1 |
| spellingShingle | Claudia Albrecht Nivethika Rajaram Julian Broche Pavel Bashtrykov Albert Jeltsch Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 Genes epigenome editing imprinting DNA demethylation dCas9 SunTag TET1 |
| title | Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 |
| title_full | Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 |
| title_fullStr | Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 |
| title_full_unstemmed | Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 |
| title_short | Locus-Specific and Stable DNA Demethylation at the <i>H19</i>/<i>IGF2</i> ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 |
| title_sort | locus specific and stable dna demethylation at the i h19 i i igf2 i icr1 by epigenome editing using a dcas9 suntag system and the catalytic domain of tet1 |
| topic | epigenome editing imprinting DNA demethylation dCas9 SunTag TET1 |
| url | https://www.mdpi.com/2073-4425/15/1/80 |
| work_keys_str_mv | AT claudiaalbrecht locusspecificandstablednademethylationattheih19iiigf2iicr1byepigenomeeditingusingadcas9suntagsystemandthecatalyticdomainoftet1 AT nivethikarajaram locusspecificandstablednademethylationattheih19iiigf2iicr1byepigenomeeditingusingadcas9suntagsystemandthecatalyticdomainoftet1 AT julianbroche locusspecificandstablednademethylationattheih19iiigf2iicr1byepigenomeeditingusingadcas9suntagsystemandthecatalyticdomainoftet1 AT pavelbashtrykov locusspecificandstablednademethylationattheih19iiigf2iicr1byepigenomeeditingusingadcas9suntagsystemandthecatalyticdomainoftet1 AT albertjeltsch locusspecificandstablednademethylationattheih19iiigf2iicr1byepigenomeeditingusingadcas9suntagsystemandthecatalyticdomainoftet1 |