%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 %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}, doi = {https://doi.org/10.1016/j.crmeth.2024.100739}, interhash = {4bf319f559120be06fb4984a99cfd1d0}, intrahash = {642dac01f1a127fd788fe27db5d9791a}, issn = {2667-2375}, journal = {Cell Reports Methods}, keywords = {}, pages = 100739, timestamp = {2024-04-11T13:56:05.000+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 }