CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci

Clustered regularly interspaced short palindromic repeats (CRISPR)-based genomic imaging systems predominantly rely on fluorescent protein reporters, which lack the optical properties essential for sensitive dynamic imaging. Here, we modified the CRISPR single-guide RNA (sgRNA) to carry two distinct...

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Detalles Bibliográficos
Autores principales: Mao, Shiqi, Ying, Yachen, Wu, Xiaotian, Krueger, Christopher J, Chen, Antony K
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6847002/
https://www.ncbi.nlm.nih.gov/pubmed/31504824
http://dx.doi.org/10.1093/nar/gkz752
Descripción
Sumario:Clustered regularly interspaced short palindromic repeats (CRISPR)-based genomic imaging systems predominantly rely on fluorescent protein reporters, which lack the optical properties essential for sensitive dynamic imaging. Here, we modified the CRISPR single-guide RNA (sgRNA) to carry two distinct molecular beacons (MBs) that can undergo fluorescence resonance energy transfer (FRET) and demonstrated that the resulting system, CRISPR/dual-FRET MB, enables dynamic imaging of non-repetitive genomic loci with only three unique sgRNAs.

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