Allele-specific DNA demethylation editing leads to stable upregulation of allele-specific gene expression

被引:1
作者
Rajaram, Nivethika [1 ]
Benzler, Katharina [1 ]
Bashtrykov, Pavel [1 ]
Jeltsch, Albert [1 ]
机构
[1] Univ Stuttgart, Inst Biochem & Tech Biochem, Allmandring 31, D-70569 Stuttgart, Germany
关键词
CATALYTIC DOMAIN; METHYLATION; CRISPR; 5-METHYLCYTOSINE; TECHNOLOGIES; SYSTEM;
D O I
10.1016/j.isci.2024.111007
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Epigenome editing is an emerging technology that allows to rewrite epigenome states and reprogram gene expression. Here, we have developed allele-specific DNA demethylation editing at gene promoters containing an SNP by sgRNA/dCas9 mediated recuitment of TET1. Maximal DNA demethylation (up to 90%) was observed 6 days after transient transfection of the epigenome editors and it was almost stable for 15 days. After allele-specific targeting, DNA demethylation was up to 15-fold more efficient at the targeted allele. Our data show that locus-specific and allele-specific DNA demethylation can trigger the expression of previously silenced genes. Allele-specific DNA demethylation shifted allelic expression ratios about 4-fold. Allele-specific DNA demethylation could be used to correct aberrant imprinting in patients suffering from imprinting disorders and to study the roles of individual alleles of a gene in a given cellular context.
引用
收藏
页数:16
相关论文
共 52 条
[1]   The Galaxy platform for accessible, reproducible, and collaborative data analyses: 2024 update [J].
Abueg, Linelle Ann L. ;
Afgan, Enis ;
Allart, Olivier ;
Wan, Ahmed H. A. ;
Bacon, W. Endi A. ;
Baker, Dannon ;
Bassetti, Madeline ;
Batut, Berenice ;
Bernt, Matthias ;
Blankenberg, Daniel ;
Bombarely, Aureliano ;
Bretaudeau, Anthony ;
Bromhead, Catherine J. ;
Burke, Melissa L. ;
Capon, Patrick K. ;
Cech, Martin ;
Chavero-Diez, Maria ;
Chilton, John M. ;
Collins, Tyler J. ;
Coppens, Frederik ;
Coraor, Nate ;
Cuccuru, Gianmauro ;
Cumbo, Fabio ;
Davis, John ;
De Geest, Paul F. ;
de Koning, Willem ;
Demko, Martin ;
DeSanto, Assunta ;
Begines, Jose Manuel Dominguez ;
Doyle, Maria A. ;
Droesbeke, Bert ;
Erxleben-Eggenhofer, Anika ;
Foll, Melanie C. ;
Formenti, Giulio ;
Fouilloux, Anne ;
Gangazhe, Rendani ;
Genthon, Tanguy ;
Goecks, Jeremy ;
Beltran, Alejandra N. Gonzalez ;
Goonasekera, Nuwan A. ;
Goue, Nadia ;
Griffin, Timothy J. ;
Gruning, Bjorn A. ;
Guerler, Aysam ;
Gundersen, Sveinung ;
Gustafsson, Ove Johan Ragnar ;
Hall, Christina ;
Harrop, Thomas W. ;
Hecht, Helge ;
Heidari, Alireza .
NUCLEIC ACIDS RESEARCH, 2024, 52 (W1) :W83-W94
[2]  
Albrecht Claudia, 2024, Methods Mol Biol, V2842, P405, DOI 10.1007/978-1-0716-4051-7_21
[3]   Locus-Specific and Stable DNA Demethylation at the H19/IGF2 ICR1 by Epigenome Editing Using a dCas9-SunTag System and the Catalytic Domain of TET1 [J].
Albrecht, Claudia ;
Rajaram, Nivethika ;
Broche, Julian ;
Bashtrykov, Pavel ;
Jeltsch, Albert .
GENES, 2024, 15 (01)
[4]   The molecular hallmarks of epigenetic control [J].
Allis, C. David ;
Jenuwein, Thomas .
NATURE REVIEWS GENETICS, 2016, 17 (08) :487-500
[5]   Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases [J].
Bae, Sangsu ;
Park, Jeongbin ;
Kim, Jin-Soo .
BIOINFORMATICS, 2014, 30 (10) :1473-1475
[6]  
Blankenberg Daniel, 2010, Curr Protoc Mol Biol, VChapter 19, DOI 10.1002/0471142727.mb1910s89
[7]   Manipulation of FASTQ data with Galaxy [J].
Blankenberg, Daniel ;
Gordon, Assaf ;
Von Kuster, Gregory ;
Coraor, Nathan ;
Taylor, James ;
Nekrutenko, Anton .
BIOINFORMATICS, 2010, 26 (14) :1783-1785
[8]   Genome-wide investigation of the dynamic changes of epigenome modifications after global DNA methylation editing [J].
Broche, Julian ;
Kungulovski, Goran ;
Bashtrykov, Pavel ;
Rathert, Philipp ;
Jeltsch, Albert .
NUCLEIC ACIDS RESEARCH, 2021, 49 (01) :158-176
[9]   Imprinting disorders in humans: a review [J].
Butler, Merlin G. .
CURRENT OPINION IN PEDIATRICS, 2020, 32 (06) :719-729
[10]   Role of Mammalian DNA Methyltransferases in Development [J].
Chen, Zhiyuan ;
Zhang, Yi .
ANNUAL REVIEW OF BIOCHEMISTRY, VOL 89, 2020, 89 :135-158