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Related Concept Videos

Experimental RNAi02:15

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 Interference01:23

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 Interference01:23

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...
siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
Small interfering RNAs (siRNA)02:30

Small interfering RNAs (siRNA)

Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...

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Related Experiment Video

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DNA Vector-based RNA Interference to Study Gene Function in Cancer
13:10

DNA Vector-based RNA Interference to Study Gene Function in Cancer

Published on: June 4, 2012

A simple and robust vector-based shRNA expression system used for RNA interference.

Xue-jun Wang1, Ying Li, Hai Huang

  • 1Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing, People's Republic of China.

Plos One
|February 14, 2013
PubMed
Summary

This study introduces a cost-effective and efficient method for creating short hairpin RNA (shRNA) expression systems. The new system enables potent gene silencing, offering a valuable tool for RNA interference (RNAi) research.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • RNA interference (RNAi) is a key genetic tool for functional studies.
  • Existing vector-based shRNA expression systems are often error-prone and expensive.

Purpose of the Study:

  • To develop a simple, robust, and cost-effective shRNA expression system.
  • To optimize shRNA design for enhanced RNA interference (RNAi) efficiency.
  • To enable rapid combination of promoter-shRNA cassettes for versatile gene silencing.

Main Methods:

  • Utilized 1 long or 2 short oligonucleotides for shRNA construction, reducing costs by 50%.
  • Optimized shRNA loop sequences and stem structures for improved RNAi efficacy.
  • Developed an isocaudomer-based strategy for rapid assembly of promoter-shRNA cassettes.
  • Screened conservative target sites for hepatitis B virus (HBV) knockdown.

Main Results:

  • >95% cloning success rate achieved with the novel shRNA system.
  • Demonstrated successful suppression of HBV antigen expression in vitro and in vivo using multiple shRNAs.
  • Identified effective promoter-shRNA cassettes for potent gene silencing.

Conclusions:

  • The developed system provides an inexpensive and effective method for cloning and expressing single or multiple shRNAs.
  • This approach allows for potent and effective silencing of target genes.
  • The system facilitates the creation of versatile gene silencing vectors.