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Nat Methods. 2018 Nov;15(11):928-931. doi: 10.1038/s41592-018-0174-0. Epub 2018 Oct 30.

CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging.

Author information

1
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA. mahh@shanghaitech.edu.cn.
2
School of Life Science and Technology, ShanghaiTech University, Shanghai, China. mahh@shanghaitech.edu.cn.
3
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA.
4
Department of Computer Science, University of Central Florida, Orlando, FL, USA.
5
Kavli Institute for the Physics and Mathematics of the Universe, University of Tokyo, Kashiwa, Japan.
6
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA.

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) guide RNA scaffolds have been adapted to carry multiple binding sites for fluorescent proteins to enhance brightness for live cell imaging of genomic loci. However, many of these modifications result in guide RNA instability and thus produce lower genome-labeling efficiency than anticipated. Here we introduce CRISPR-Sirius, based on octet arrays of aptamers conferring both enhanced guide RNA stability and brightness, and provide initial biological applications of this platform.

PMID:
30377374
PMCID:
PMC6252086
DOI:
10.1038/s41592-018-0174-0
[Indexed for MEDLINE]
Free PMC Article

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