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Designer Receptors for Nucleotide‐Resolution Analysis of Genomic 5‐Methylcytosine by Cellular Imaging by Álvaro Muñoz‐López; Benjamin Buchmuller; Jan Wolffgramm; Anne Jung; Michelle Hussong; Julian Kanne; Michal R. Schweiger; Daniel Summerer is a Chemistry article available to read on EtoBox.

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## 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.

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Author

Álvaro Muñoz‐López; Benjamin Buchmuller; Jan Wolffgramm; Anne Jung; Michelle Hussong; Julian Kanne; Michal R. Schweiger; Daniel Summerer

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley; Research Square (ISSN 0044-8249)

Published

2020

Language

EN

Field

Chemistry (Physical Sciences)