CRISPR/Cas9-Based RGEN-ISL Allows the Simultaneous and Specific Visualization of Proteins, DNA Repeats, and Sites of DNA Replication
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61389030%3A_____%2F19%3A00517551" target="_blank" >RIV/61389030:_____/19:00517551 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1159/000502600" target="_blank" >http://dx.doi.org/10.1159/000502600</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1159/000502600" target="_blank" >10.1159/000502600</a>
Alternative languages
Result language
angličtina
Original language name
CRISPR/Cas9-Based RGEN-ISL Allows the Simultaneous and Specific Visualization of Proteins, DNA Repeats, and Sites of DNA Replication
Original language description
Visualizing the spatiotemporal organization of the genome will improve our understanding of how chromatin structure and function are intertwined. Here, we describe a further development of the CRISPR/Cas9-based RNA-guided endonuclease-in situ labeling (RGEN-ISL) method. RGEN-ISL allowed the differentiation between vertebrate-type (TTAGGG)n and Arabidopsis-type (TTTAGGG)n telomere repeats. Using maize as an example, we established a combination of RGEN-ISL, immunostaining, and EdU labeling to visualize in situ specific repeats, histone marks, and DNA replication sites, respectively. The effects of the non-denaturing RGEN-ISL and standard denaturing FISH on the chromatin structure were compared using super-resolution microscopy. 3D structured illumination microscopy revealed that denaturation and acetic acid fixation impaired and flattened the chromatin. The broad range of adaptability of RGEN-ISL to different combinations of methods has the potential to advance the field of chromosome biology.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10603 - Genetics and heredity (medical genetics to be 3)
Result continuities
Project
<a href="/en/project/GA17-14048S" target="_blank" >GA17-14048S: Spatial and temporal characterization of DNA replication in phylogeneticaly related plant species with contrasting genome sizes</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2019
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Cytogenetic and Genome Research
ISSN
1424-8581
e-ISSN
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Volume of the periodical
159
Issue of the periodical within the volume
1
Country of publishing house
CH - SWITZERLAND
Number of pages
6
Pages from-to
48-53
UT code for WoS article
000498640000007
EID of the result in the Scopus database
2-s2.0-85074473340