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Nat Cell Biol ; 23(1): 99-108, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33398178

RESUMO

Detection of endogenous signals and precise control of genetic circuits in the natural context are essential to understand biological processes. However, the tools to process endogenous information are limited. Here we developed a generalizable endogenous transcription-gated switch that releases single-guide RNAs in the presence of an endogenous promoter. When the endogenous transcription-gated switch is coupled with the highly sensitive CRISPR-activator-associated reporter we developed, we can reliably detect the activity of endogenous genes, including genes with very low expression (<0.001 relative to Gapdh; quantitative-PCR analysis). Notably, we could also monitor the transcriptional activity of typically long non-coding RNAs expressed at low levels in living cells using this approach. Together, our method provides a powerful platform to sense the activity of endogenous genetic elements underlying cellular functions.


Assuntos
Células-Tronco Embrionárias Murinas/metabolismo , Neuroblastoma/genética , Neuroblastoma/metabolismo , Regiões Promotoras Genéticas , RNA Guia de Cinetoplastídeos/metabolismo , RNA Longo não Codificante/metabolismo , RNA Mensageiro/metabolismo , Animais , Sistemas CRISPR-Cas , Células HEK293 , Humanos , Camundongos , Células-Tronco Embrionárias Murinas/citologia , Neuroblastoma/patologia , RNA Guia de Cinetoplastídeos/genética , RNA Longo não Codificante/genética , RNA Mensageiro/genética
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