Universität zu Köln

Munoz-Lopez, Alvaro, Buchmuller, Benjamin ORCID: 0000-0002-4915-5949, Wolffgramm, Jan, Jung, Anne, Hussong, Michelle, Kanne, Julian, Schweiger, Michal R. and Summerer, Daniel (2020). Designer Receptors for Nucleotide-Resolution Analysis of Genomic 5-Methylcytosine by Cellular Imaging. Angew. Chem.-Int. Edit., 59 (23). S. 8927 - 8932. WEINHEIM: WILEY-V C H VERLAG GMBH. ISSN 1521-3773

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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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Munoz-Lopez, Alvaro UNSPECIFIED UNSPECIFIED UNSPECIFIED Buchmuller, Benjamin UNSPECIFIED orcid.org/0000-0002-4915-5949 UNSPECIFIED Wolffgramm, Jan UNSPECIFIED UNSPECIFIED UNSPECIFIED Jung, Anne UNSPECIFIED UNSPECIFIED UNSPECIFIED Hussong, Michelle UNSPECIFIED UNSPECIFIED UNSPECIFIED Kanne, Julian UNSPECIFIED UNSPECIFIED UNSPECIFIED Schweiger, Michal R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Summerer, Daniel UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-337476
DOI:	10.1002/anie.202001935
Journal or Publication Title:	Angew. Chem.-Int. Edit.
Volume:	59
Number:	23
Page Range:	S. 8927 - 8932
Date:	2020
Publisher:	WILEY-V C H VERLAG GMBH
Place of Publication:	WEINHEIM
ISSN:	1521-3773
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DNA METHYLATION; TRANSCRIPTION; RECOGNITION; SEQUENCES; VISUALIZATION; ORGANIZATION; REPEATS; PROTEIN; CELLS Multiple languages Chemistry, Multidisciplinary Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/33747