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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...

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...

Types of RNA

Types of RNA

Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...

Types of RNA

Types of RNA

Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in regulating gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA Performs Diverse...

Translational Regulation

Translational Regulation

Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...

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Related Experiment Video

Updated: May 27, 2026

Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells

Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells

Published on: April 25, 2018

[shRNAs driven by K14 promoter induce tissue-specific RNA interference].

Rong Dai¹, Si-Jun Shen, Peng-Cheng Wan

¹College of Animal Science, Shihezi University, Shihezi 832000, China. dairong1@163.com

Yi Chuan = Hereditas

|November 5, 2011

Summary

RNA interference effectively silences gene expression using RNA Pol II promoters. This study demonstrates efficient BMP4 gene silencing in vitro and in vivo, offering a novel RNAi application for hair follicle development research.

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Bacterial Delivery of RNAi Effectors: Transkingdom RNAi

Bacterial Delivery of RNAi Effectors: Transkingdom RNAi

Published on: August 18, 2010

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Last Updated: May 27, 2026

Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells

Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells

Published on: April 25, 2018

Bacterial Delivery of RNAi Effectors: Transkingdom RNAi

Bacterial Delivery of RNAi Effectors: Transkingdom RNAi

Published on: August 18, 2010

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

RNA interference (RNAi) is a powerful tool for gene function analysis.
RNA Pol II promoters can mediate cell- and tissue-specific gene silencing.
BMP4 gene expression plays a role in biological processes.

Purpose of the Study:

To develop and evaluate an RNA Pol II promoter-driven shRNA system for BMP4 gene silencing.
To assess the efficiency of gene-specific shRNA in both in vitro and in vivo models.
To explore the potential application of this RNAi system in hair follicle development research.

Main Methods:

Construction of recombinant vectors expressing eGFP-shRNA fusion constructs under the K14 promoter.
In vitro cotransfection of Hela cells with vectors and assessment of BMP4 silencing using luciferase assays.
In vivo screening in JB6-C41 cells using quantitative RT-PCR and Western blotting to evaluate gene silencing efficiency.

Main Results:

All tested shRNA sequences demonstrated high efficiency (>60%) in inhibiting BMP4 expression in vitro.
Sequence 5# exhibited the highest inhibition efficiency in vitro.
In vivo, the K14 promoter-driven shRNA system efficiently and specifically inhibited both BMP4 mRNA and protein levels (>60% efficiency, except for sequence 3#).
A significant correlation was observed between the reduction in mRNA and protein levels during gene silencing.

Conclusions:

The K14 promoter can effectively drive shRNA expression for BMP4 gene silencing.
The developed RNAi system shows high efficiency and specificity for gene silencing in vitro and in vivo.
This approach offers a promising new method for applying RNAi technology to study gene expression mechanisms, such as those in sheep hair follicle development.