Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F21%3A00119668" target="_blank" >RIV/00216224:14740/21:00119668 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00159816:_____/21:00075086 RIV/00216208:11310/21:10442523

Výsledek na webu

<a href="https://journals.biologists.com/jcs/article/134/15/jcs258374/271206/Super-resolution-microscopy-of-chromatin-fibers" target="_blank" >https://journals.biologists.com/jcs/article/134/15/jcs258374/271206/Super-resolution-microscopy-of-chromatin-fibers</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1242/jcs.258374" target="_blank" >10.1242/jcs.258374</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations

Popis výsledku v původním jazyce

Analysis of histone variants and epigenetic marks is dominated by genome-wide approaches in the form of chromatin immunoprecipitation-sequencing (ChIP-seq) and related methods. Although uncontested in their value for single-copy genes, mapping the chromatin of DNA repeats is problematic for biochemical techniques that involve averaging of cell populations or analysis of clusters of tandem repeats in a single-cell analysis. Extending chromatin and DNA fibers allows us to study the epigenetics of individual repeats in their specific chromosomal context, and thus constitutes an important tool for gaining a complete understanding of the epigenetic organization of genomes. We report that using an optimized fiber extension protocol is essential in order to obtain more reproducible data and to minimize the clustering of fibers. We also demonstrate that the use of super-resolution microscopy is important for reliable evaluation of the distribution of histone modifications on individual fibers. Furthermore, we introduce a custom script for the analysis of methylation levels on DNA fibers and apply it to map the methylation of telomeres, ribosomal genes and centromeres.

Název v anglickém jazyce

Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations

Popis výsledku anglicky

Analysis of histone variants and epigenetic marks is dominated by genome-wide approaches in the form of chromatin immunoprecipitation-sequencing (ChIP-seq) and related methods. Although uncontested in their value for single-copy genes, mapping the chromatin of DNA repeats is problematic for biochemical techniques that involve averaging of cell populations or analysis of clusters of tandem repeats in a single-cell analysis. Extending chromatin and DNA fibers allows us to study the epigenetics of individual repeats in their specific chromosomal context, and thus constitutes an important tool for gaining a complete understanding of the epigenetic organization of genomes. We report that using an optimized fiber extension protocol is essential in order to obtain more reproducible data and to minimize the clustering of fibers. We also demonstrate that the use of super-resolution microscopy is important for reliable evaluation of the distribution of histone modifications on individual fibers. Furthermore, we introduce a custom script for the analysis of methylation levels on DNA fibers and apply it to map the methylation of telomeres, ribosomal genes and centromeres.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10601 - Cell biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Cell Science

ISSN

0021-9533

e-ISSN

1477-9137

Svazek periodika

134

Číslo periodika v rámci svazku

15

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

14

Strana od-do

1-14

Kód UT WoS článku

000686349300013

EID výsledku v databázi Scopus

2-s2.0-85113376383