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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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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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Experimental RNAi02:15

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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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siRNA - Small Interfering RNAs02:30

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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.
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mirMachine: A One-Stop Shop for Plant miRNA Annotation
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Manipulating MiRNA Expression to Uncover Hidden Functions.

Sinéad T Aherne1,2, Nga T Lao3

  • 1Department of Tumor Biology, H. Lee Moffitt Cancer Center and Research Institute, SRB-3, 12902 Magnolia Drive, Tampa, FL, 33612, USA. Sinead.Aherne@moffitt.org.

Methods in Molecular Biology (Clifton, N.J.)
|November 10, 2016
PubMed
Summary

Discovering microRNA (miRNA) function requires more than prediction algorithms. This study details methods for physically altering miRNA expression to definitively determine their roles in biological systems.

Keywords:
CloningLigationTransfectionmiRNA-Target

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNA (miRNA) target prediction algorithms offer insights into potential regulatory roles.
  • However, these computational predictions require experimental validation for definitive functional determination.

Purpose of the Study:

  • To outline experimental methods for elucidating microRNA (miRNA) function.
  • To provide a guide for manipulating miRNA expression to study their biological roles.

Main Methods:

  • Transient transfection of miRNA mimics and inhibitors.
  • Stable gene expression modulation using plasmid overexpression vectors.
  • Utilizing sponge vectors for stable miRNA expression reduction.

Main Results:

  • Demonstration of effective miRNA expression modulation through described techniques.
  • Validation of experimental approaches for functional miRNA studies.

Conclusions:

  • Physical manipulation of miRNA expression is essential for accurate functional analysis.
  • The described methods provide a robust framework for investigating miRNA functions in various biological contexts.