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Experimental RNAi

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

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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Related Experiment Video

Updated: Jun 10, 2026

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Published on: July 16, 2011

Establishing efficient siRNA knockdown in stem cells using fluorescent oligonucleotides.

Stephen W Chen¹, Steve K W Oh

¹Stem Cell Group, Bioprocessing Technology Institute, Centros, Singapore.

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

|August 6, 2010

Summary

This study presents rapid protocols for effective gene knockdown using small interfering RNAs (siRNAs). Evaluating transfection efficiency via microscopy before costly assays optimizes experimental success.

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MISSION esiRNA for RNAi Screening in Mammalian Cells

MISSION esiRNA for RNAi Screening in Mammalian Cells

Published on: May 12, 2010

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Last Updated: Jun 10, 2026

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Published on: July 16, 2011

MISSION esiRNA for RNAi Screening in Mammalian Cells

MISSION esiRNA for RNAi Screening in Mammalian Cells

Published on: May 12, 2010

Area of Science:

Molecular Biology
Cell Biology
Biotechnology

Background:

Gene knockdown experiments using small interfering RNAs (siRNAs) are crucial for studying gene function.
Optimizing transfection efficiency is a common challenge in siRNA-based gene silencing.
Ineffective knockdown is often attributed to poor cellular uptake of siRNAs.

Purpose of the Study:

To provide rapid protocols for optimizing siRNA transfection conditions.
To establish a reliable method for evaluating transfection efficiency prior to downstream analyses.
To minimize resource expenditure on troubleshooting ineffective gene knockdown experiments.

Main Methods:

Utilizing microscopy to assess the transfection efficiency of an inexpensive fluorescent oligonucleotide.
Implementing protocols for siRNA delivery and optimization.
Employing Western blot and quantitative real-time PCR (qRT-PCR) for validation.

Main Results:

Microscopy-based evaluation of fluorescent oligonucleotide uptake provides a rapid assessment of transfection efficiency.
This preliminary step effectively identifies and eliminates poor transfection as a cause of failed gene knockdown.
Successful optimization allows for more reliable downstream gene silencing validation.

Conclusions:

A microscopy-based approach offers a rapid and cost-effective method to optimize siRNA transfection.
This strategy streamlines the experimental workflow for gene knockdown studies.
Researchers can confidently proceed with validation assays, saving time and resources.