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Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Related Experiment Video

Updated: Mar 20, 2026

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases

Published on: August 16, 2024

An RNA-aptamer-based two-color CRISPR labeling system.

Siyuan Wang^1,2, Jun-Han Su^1,2, Feng Zhang^3,4,5

¹Howard Hughes Medical Institute, Cambridge, MA 02138, USA.

Scientific Reports

|May 28, 2016

Summary

This summary is machine-generated.

This study introduces a novel two-color CRISPR labeling technique for visualizing chromatin in living cells. This method enhances imaging by using engineered single-guide RNAs (sgRNAs) with RNA aptamers to recruit fluorescent proteins, improving signal clarity.

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Area of Science:

Molecular Biology
Cell Biology
Genetics

Background:

Chromatin organization and dynamics are crucial for cellular functions.
Visualizing genomic loci in living cells has been challenging.
CRISPR-Cas9 systems enable targeted chromatin labeling.

Purpose of the Study:

To develop an alternative two-color CRISPR labeling method.
To enable visualization of endogenous genomic loci in living cells.
To improve the signal-to-background ratio for chromatin imaging.

Main Methods:

Utilized Streptococcus pyogenes Cas9.
Incorporated MS2 or PP7 RNA aptamers into single-guide RNAs (sgRNAs).
Recruited MS2 or PP7 coat proteins fused with fluorescent proteins to target loci.

Main Results:

Achieved specific and orthogonal two-color labeling of repetitive sequences in human cells.
Demonstrated that aptamer attachment location influences signal-to-background ratio.
Enhanced signal-to-background ratio by extending tetraloop and stem loop 2 of sgRNA.

Conclusions:

The developed method offers a robust alternative for multicolor chromatin labeling.
This technique simplifies multicolor imaging by using a single Cas9 ortholog.
Optimized sgRNA designs improve the efficiency and clarity of chromatin visualization.