Summary
Dynamic changes in the epigenome at defined genomic loci play crucial roles during cellular differentiation and disease development. Here, we developed dual-color bimolecular anchor detector (BiAD) sensors for high-sensitivity readout of locus-specific epigenome modifications by fluorescence microscopy. Our BiAD sensors comprise an sgRNA/dCas9 complex as anchor and double chromatin reader domains as detector modules, both fused to complementary parts of a split IFP2.0 fluorophore, enabling its reconstitution upon binding of both parts in close proximity. In addition, a YPet fluorophore is recruited to the sgRNA to mark the genomic locus of interest. With these dual-color BiAD sensors, we detected H3K9me2/3 and DNA methylation and their dynamic changes upon RNAi or inhibitor treatment with high sensitivity at endogenous genomic regions. Furthermore, we showcased locus-specific H3K36me2/3 readout as well as H3K27me3 and H3K9me2/3 enrichment on the inactive X chromosome, highlighting the broad applicability of our dual-color BiAD sensors for single-cell epigenome studies.
Description
Modular dual-color BiAD sensors for locus-specific readout of epigenome modifications in single cells - ScienceDirect
%0 Journal Article
%1 KOHLER2024100739
%A Köhler, Anja R.
%A Haußer, Johannes
%A Harsch, Annika
%A Bernhardt, Steffen
%A Häußermann, Lilia
%A Brenner, Lisa-Marie
%A Lungu, Cristiana
%A Olayioye, Monilola A.
%A Bashtrykov, Pavel
%A Jeltsch, Albert
%D 2024
%J Cell Reports Methods
%K ajeltsch ibtb-bc imported myown unibiblio unibibliografie
%P 100739
%R https://doi.org/10.1016/j.crmeth.2024.100739
%T Modular dual-color BiAD sensors for locus-specific readout of epigenome modifications in single cells
%U https://www.sciencedirect.com/science/article/pii/S266723752400064X
%X Summary
Dynamic changes in the epigenome at defined genomic loci play crucial roles during cellular differentiation and disease development. Here, we developed dual-color bimolecular anchor detector (BiAD) sensors for high-sensitivity readout of locus-specific epigenome modifications by fluorescence microscopy. Our BiAD sensors comprise an sgRNA/dCas9 complex as anchor and double chromatin reader domains as detector modules, both fused to complementary parts of a split IFP2.0 fluorophore, enabling its reconstitution upon binding of both parts in close proximity. In addition, a YPet fluorophore is recruited to the sgRNA to mark the genomic locus of interest. With these dual-color BiAD sensors, we detected H3K9me2/3 and DNA methylation and their dynamic changes upon RNAi or inhibitor treatment with high sensitivity at endogenous genomic regions. Furthermore, we showcased locus-specific H3K36me2/3 readout as well as H3K27me3 and H3K9me2/3 enrichment on the inactive X chromosome, highlighting the broad applicability of our dual-color BiAD sensors for single-cell epigenome studies.
@article{KOHLER2024100739,
abstract = {Summary
Dynamic changes in the epigenome at defined genomic loci play crucial roles during cellular differentiation and disease development. Here, we developed dual-color bimolecular anchor detector (BiAD) sensors for high-sensitivity readout of locus-specific epigenome modifications by fluorescence microscopy. Our BiAD sensors comprise an sgRNA/dCas9 complex as anchor and double chromatin reader domains as detector modules, both fused to complementary parts of a split IFP2.0 fluorophore, enabling its reconstitution upon binding of both parts in close proximity. In addition, a YPet fluorophore is recruited to the sgRNA to mark the genomic locus of interest. With these dual-color BiAD sensors, we detected H3K9me2/3 and DNA methylation and their dynamic changes upon RNAi or inhibitor treatment with high sensitivity at endogenous genomic regions. Furthermore, we showcased locus-specific H3K36me2/3 readout as well as H3K27me3 and H3K9me2/3 enrichment on the inactive X chromosome, highlighting the broad applicability of our dual-color BiAD sensors for single-cell epigenome studies.},
added-at = {2024-04-11T15:56:05.000+0200},
author = {Köhler, Anja R. and Haußer, Johannes and Harsch, Annika and Bernhardt, Steffen and Häußermann, Lilia and Brenner, Lisa-Marie and Lungu, Cristiana and Olayioye, Monilola A. and Bashtrykov, Pavel and Jeltsch, Albert},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2642dac01f1a127fd788fe27db5d9791a/ajeltsch},
description = {Modular dual-color BiAD sensors for locus-specific readout of epigenome modifications in single cells - ScienceDirect},
doi = {https://doi.org/10.1016/j.crmeth.2024.100739},
interhash = {4bf319f559120be06fb4984a99cfd1d0},
intrahash = {642dac01f1a127fd788fe27db5d9791a},
issn = {2667-2375},
journal = {Cell Reports Methods},
keywords = {ajeltsch ibtb-bc imported myown unibiblio unibibliografie},
pages = 100739,
timestamp = {2024-04-11T15:56:05.000+0200},
title = {Modular dual-color BiAD sensors for locus-specific readout of epigenome modifications in single cells},
url = {https://www.sciencedirect.com/science/article/pii/S266723752400064X},
year = 2024
}