Transcription-coupled changes in nuclear mobility of mammalian cis-regulatory elements.
Science
; 359(6379): 1050-1055, 2018 03 02.
Article
em En
| MEDLINE
| ID: mdl-29371426
ABSTRACT
To achieve guide RNA (gRNA) multiplexing and an efficient delivery of tens of distinct gRNAs into single cells, we developed a molecular assembly strategy termed chimeric array of gRNA oligonucleotides (CARGO). We coupled CARGO with dCas9 (catalytically dead Cas9) imaging to quantitatively measure the movement of enhancers and promoters that undergo differentiation-associated activity changes in live embryonic stem cells. Whereas all examined functional elements exhibited subdiffusive behavior, their relative mobility increased concurrently with transcriptional activation. Furthermore, acute perturbation of RNA polymerase II activity can reverse these activity-linked increases in loci mobility. Through quantitative CARGO-dCas9 imaging, we provide direct measurements of cis-regulatory element dynamics in living cells and distinct cellular and activity states and uncover an intrinsic connection between cis-regulatory element mobility and transcription.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Transcrição Gênica
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Sequências Reguladoras de Ácido Nucleico
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RNA Guia de Cinetoplastídeos
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Análise de Célula Única
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Imagem Individual de Molécula
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article