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
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author Mao, Shiqi
Ying, Yachen
Wu, Xiaotian
Krueger, Christopher J
Chen, Antony K
author_facet Mao, Shiqi
Ying, Yachen
Wu, Xiaotian
Krueger, Christopher J
Chen, Antony K
author_sort Mao, Shiqi
collection PubMed
description 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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spelling pubmed-68470022019-11-18 CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci Mao, Shiqi Ying, Yachen Wu, Xiaotian Krueger, Christopher J Chen, Antony K Nucleic Acids Res Methods Online 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. Oxford University Press 2019-11-18 2019-08-31 /pmc/articles/PMC6847002/ /pubmed/31504824 http://dx.doi.org/10.1093/nar/gkz752 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Mao, Shiqi
Ying, Yachen
Wu, Xiaotian
Krueger, Christopher J
Chen, Antony K
CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
title CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
title_full CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
title_fullStr CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
title_full_unstemmed CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
title_short CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
title_sort crispr/dual-fret molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci
topic Methods Online
url 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
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AT kruegerchristopherj crisprdualfretmolecularbeaconforsensitivelivecellimagingofnonrepetitivegenomicloci
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