Determination of local chromatin interactions using a combined CRISPR and peroxidase APEX2 system.
Nucleic Acids Res
; 47(9): e52, 2019 05 21.
Article
em En
| MEDLINE
| ID: mdl-30805613
ABSTRACT
The architecture and function of chromatin are largely regulated by local interacting molecules, such as transcription factors and noncoding RNAs. However, our understanding of these regulatory molecules at a given locus is limited because of technical difficulties. Here, we describe the use of Clustered Regularly Interspaced Short Palindromic Repeats and an engineered ascorbate peroxidase 2 (APEX2) system to investigate local chromatin interactions (CAPLOCUS). We showed that with specific small-guide RNA targets, CAPLOCUS could efficiently identify both repetitive genomic regions and single-copy genomic locus with high resolution. Genome-wide sequencing revealed known and potential long-range chromatin interactions for a specific single-copy locus. CAPLOCUS also identified telomere-associated RNAs. CAPLOCUS, followed by mass spectrometry, identified both known and novel telomere-associated proteins in their native states. Thus, CAPLOCUS may be a useful approach for studying local interacting molecules at any given chromosomal location.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Cromatina
/
Sequências Repetitivas de Ácido Nucleico
/
RNA Guia de Cinetoplastídeos
/
DNA Liase (Sítios Apurínicos ou Apirimidínicos)
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Nucleic Acids Res
Ano de publicação:
2019
Tipo de documento:
Article