Authors: Muñoz-López, Álvaro
Buchmuller, Benjamin
Wolffgramm, Jan
Jung, Anne
Hussong, Michelle
Kanne, Julian
Schweiger, Michal Ruth
Summerer, Daniel
Title: Designer receptors for nucleotide-resolution analysis of genomic 5-Methylcytosine by cellular imaging
Language (ISO): en
Abstract: We report programmable receptors for the imaging-based analysis of 5-methylcytosine (5mC) in user-defined DNA sequences of single cells. Using fluorescent transcription-activator-like effectors (TALEs) that can recognize sequences of canonical and epigenetic nucleobases through selective repeats, we imaged cellular SATIII DNA, the origin of nuclear stress bodies (nSB). We achieve high nucleobase selectivity of natural repeats in imaging and demonstrate universal nucleobase binding by an engineered repeat. We use TALE pairs differing in only one such repeat in co-stains to detect 5mC in SATIII sequences with nucleotide resolution independently of differences in target accessibility. Further, we directly correlate the presence of heat shock factor 1 with 5mC at its recognition sequence, revealing a potential function of 5mC in its recruitment as initial step of nSB formation. This opens a new avenue for studying 5mC functions in chromatin regulation in situ with nucleotide, locus, and cell resolution.
Subject Headings: Biosensors
DNA methylation
Imaging probes
Membrane-less organelles
Subject Headings (RSWK): Biosensor
Issue Date: 2020-03-13
Appears in Collections:Chemische Biologie

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