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Related Concept Videos

siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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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...
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RNA Interference01:23

RNA Interference

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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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Experimental RNAi02:15

Experimental RNAi

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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...
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Types of RNA01:20

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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...
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MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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Related Experiment Video

Updated: Apr 14, 2026

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
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Targeting Human Long Noncoding Transcripts by Endoribonuclease-Prepared siRNAs.

Mirko Theis1, Maciej Paszkowski-Rogacz2, Ina Weisswange3

  • 1Medical Systems Biology, UCC, Medical Faculty Carl Gustav Carus, TU Dresden, Dresden, Germany Eupheria Biotech GmbH, Dresden, Germany mirko.theis@tu-dresden.de.

Journal of Biomolecular Screening
|April 26, 2015
PubMed
Summary

Researchers developed a new tool to study long noncoding RNAs (lncRNAs). This endoribonuclease-prepared siRNA library enables cost-effective functional analysis of these important, yet poorly understood, RNA molecules.

Keywords:
RNAiesiRNAlncRNAlocalizationnoncoding RNA

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Area of Science:

  • Genomics
  • Molecular Biology
  • Transcriptomics

Background:

  • Large-scale sequencing projects like FANTOM and ENCODE have revealed extensive transcription of the human genome.
  • A significant portion of these transcripts are long noncoding RNAs (lncRNAs) with largely unknown functions.
  • Efficient tools are needed to investigate the biological roles of the vast number of identified lncRNAs.

Purpose of the Study:

  • To create and validate a resource for functional analysis of human long noncoding RNAs (lncRNAs).
  • To provide a compendium of lncRNA expression in human cancer cell lines.
  • To facilitate combined knockdown and localization studies for lncRNAs.

Main Methods:

  • Construction and validation of an endoribonuclease-prepared siRNA (esiRNA) library targeting 1779 human lncRNAs.
  • RNA interference (RNAi) based knockdown of lncRNAs.
  • Analysis of lncRNA expression profiles across 11 human cancer cell lines.
  • Assessment of esiRNA suitability for knockdown and cellular localization studies.

Main Results:

  • A comprehensive library of 1779 human lncRNA-targeting esiRNAs was successfully constructed and validated.
  • A dataset of lncRNA expression was generated for 11 human cancer cell lines.
  • The esiRNA library demonstrated efficacy in combined knockdown and localization analyses.
  • Challenges related to lncRNA annotation, including cell-type-specific expression, were highlighted.

Conclusions:

  • The developed esiRNA library provides a valuable and cost-effective tool for functional lncRNA research.
  • The lncRNA expression compendium offers insights into cancer-related transcriptomes.
  • This resource supports further investigation into the functions of lncRNAs in various cellular contexts.