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RNA Interference

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...

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Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface

Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface

Published on: November 2, 2011

RNA Interference Approaches to Study Epidermal Cell Adhesion.

Anamika Dutta¹, Michele Calder¹, Lina Dagnino^2,3

¹Department of Physiology and Pharmacology, University of Western Ontario, London, ON, Canada.

Methods in Molecular Biology (Clifton, N.J.)

|January 17, 2025

Summary

This chapter details a method for gene silencing in epidermal cells using RNA interference, specifically through small interfering RNA (siRNA) transfection. It also presents assays to study how gene knockdown affects cell adhesion properties.

Keywords:

Cell adhesion Epidermal squamous carcinoma cells Gene silencing Keratinocyte RNA interference siRNA transfection

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Area of Science:

Molecular Biology
Cell Biology
Genetics

Background:

Gene silencing is crucial for understanding gene function.
Epidermal cells play vital roles in tissue integrity and barrier function.
RNA interference (RNAi) offers a powerful tool for targeted gene knockdown.

Purpose of the Study:

To provide a detailed protocol for gene silencing in epidermal cells using RNA interference.
To describe the methodology for transfecting small interfering RNA (siRNA) oligonucleotides.
To present functional assays for analyzing the consequences of gene knockdown on cell adhesion.

Main Methods:

RNA interference (RNAi) for gene silencing.
Transfection-mediated delivery of small interfering RNA (siRNA) oligonucleotides.
Functional assays to assess cell adhesion properties post-gene knockdown.

Main Results:

Successful implementation of a protocol for siRNA-mediated gene silencing in epidermal cells.
Demonstration of functional assays to evaluate changes in cell adhesion.
Characterization of biological consequences resulting from specific gene knockdowns.

Conclusions:

The described method enables effective gene silencing in epidermal cells.
The provided assays facilitate the study of gene function, particularly in relation to cell adhesion.
This chapter serves as a valuable resource for researchers investigating gene function in epidermal biology.