Related Experiment Video
Updated: Feb 8, 2026
12:26
Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation
Published on: February 12, 2022
5.9K
Single-Cell Imaging Approaches for Studying Small-RNA-Induced Gene Regulation
1T.C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, Maryland; Department of Chemistry, Chung-Ang University, Seoul, Korea.
Biophysical Journal
|July 5, 2018
Summary
We developed a single-cell mRNA imaging technique to precisely measure RNA interference (RNAi) efficiency. This method visualizes individual messenger RNA targets, revealing cellular localization and heterogeneity for potent small interfering RNA or microRNA assessment.
Area of Science:
- Molecular Biology
- Biophysics
- Genetics
Background:
- RNA interference (RNAi) is a crucial gene regulatory process.
- Quantifying RNAi efficiency at messenger RNA (mRNA) and protein levels is essential for assessing small interfering RNA (siRNA) and microRNA (miRNA) potency.
- Existing methods may not fully capture cellular heterogeneity or localization dynamics.
Purpose of the Study:
- To introduce and highlight a novel single-cell mRNA imaging method for quantitative analysis of RNAi.
- To demonstrate the visualization of individual mRNA targets with high precision.
- To discuss future directions and applicable single-cell imaging techniques for RNAi research.
Main Methods:
- Single-cell mRNA imaging techniques were employed.
- Visualization of individual mRNA targets at high precision.
- Quantitative analysis of RNAi efficiency, cellular localization, and cell-to-cell heterogeneity.
Main Results:
- The study successfully visualized individual mRNA targets in real-time.
- High precision in quantifying RNAi efficiency was achieved.
- Cellular localization and heterogeneity of RNAi effects were resolved.
Conclusions:
- Single-cell mRNA imaging provides a powerful tool for quantitative RNAi analysis.
- This technique offers insights into cellular localization and heterogeneity of gene silencing.
- The discussed imaging methods are broadly applicable to other gene regulation processes, including CRISPR-Cas systems.
Related Concept Videos
01:17Gene Regulation During Sporulation
515
Sporulation is a complex developmental process that allows certain Gram-positive bacteria, such as Bacillus subtilis and Clostridium species, to survive extreme environmental conditions. This process is tightly regulated by a series of signaling cascades and transcriptional controls, ensuring the formation of a highly resistant endospore.Sporulation is triggered by unfavorable conditions, such as nutrient depletion, and is governed by a phosphorelay system. One of the sensor kinases, such as...
515
01:58Cell Specific Gene Expression
16.6K
Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
16.6K
01:46Epigenetic Regulation
33.9K
Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
33.9K
02:18Alternative RNA Splicing
25.2K
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
25.2K
02:19Circadian Rhythms and Gene Regulation
4.6K
The biological clock is involved in many aspects of regulating complex physiology in all animals. It was in 1935 when German zoologists, Hans Kalmus and Erwin Bünning, discovered the existence of circadian rhythm in Drosophila melanogaster. However, the internal molecular mechanisms behind the circadian clock remained a mystery until 1984, when Jeffrey C. Hall, Michael Rosbash, and Michael W. Young discovered the expression of the Per gene oscillating over a 24-hour cycle. In subsequent...
4.6K
01:27Constitutive and Regulated Gene Expression
1.2K
Gene expression in prokaryotes is governed by constitutive and regulated systems, allowing cells to balance the production of essential proteins with adaptive responses to environmental changes.Constitutive Gene ExpressionConstitutive, or housekeeping, genes are continuously expressed as they encode proteins vital for fundamental cellular processes. These include enzymes for glycolysis, ribosomal components for protein synthesis, and proteins involved in DNA replication. Their constant...
1.2K