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

Shiqi Mao; Yachen Ying; Xiaotian Wu; Christopher J Krueger; Antony K Chen

doi:10.1093/nar/gkz752

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

Nucleic Acids Res. 2019 Nov 18;47(20):e131. doi: 10.1093/nar/gkz752.

Authors

Shiqi Mao¹, Yachen Ying¹, Xiaotian Wu^{1

2}, Christopher J Krueger^{1

3}, Antony K Chen¹

Affiliations

¹ Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China.
² School of Life Sciences, Peking University, Beijing 100871, China.
³ Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems*
Fluorescence Resonance Energy Transfer / methods*
Fluorescent Dyes / chemistry
Genetic Loci*
HeLa Cells
Humans
RNA, Guide, CRISPR-Cas Systems / chemistry
RNA, Guide, CRISPR-Cas Systems / metabolism

Substances

Fluorescent Dyes
RNA, Guide, CRISPR-Cas Systems