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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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...
MicroRNAs01:22

MicroRNAs

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 ends...
MicroRNAs01:22

MicroRNAs

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 ends...
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...
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...

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Related Experiment Video

Updated: May 23, 2026

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
06:48

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

Published on: June 16, 2022

Precursor microRNA-programmed silencing complex assembly pathways in mammals.

Xuhang Liu1, Dong-Yan Jin, Michael T McManus

  • 1Department of Pathology and Laboratory Medicine, Division of Neuropathology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.

Molecular Cell
|April 17, 2012
PubMed
Summary
This summary is machine-generated.

This study reveals key mechanisms of microRNA (miRNA) ribonucleoprotein (miRNP) assembly in mammals. A novel in vitro assay and in vivo studies identify the miRNA loading complex and miRNA precursor deposit complex as critical for miRNA biogenesis.

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Last Updated: May 23, 2026

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Published on: June 16, 2022

mirMachine: A One-Stop Shop for Plant miRNA Annotation
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11:00

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

Published on: June 12, 2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNA (miRNA) ribonucleoproteins (miRNPs) and RNA-induced silencing complexes (RISCs) are crucial for gene regulation.
  • miRNPs/RISCs assemble from precursor miRNAs (pre-miRNAs) processed by Dicer or Ago2.

Purpose of the Study:

  • To investigate the in vitro assembly of miRNPs/RISCs using pre-miRNAs in mammalian cell lysates.
  • To elucidate the roles of the miRNA loading complex (miRLC) and miRNA precursor deposit complex (miPDC) in miRNA biogenesis and assembly.

Main Methods:

  • Development of an in vitro miRNP/RISC assembly assay using pre-miRNAs.
  • In vivo studies utilizing Dicer knockout cells reconstituted with wild-type or catalytically inactive Dicer.
  • Analysis of pre-miRNA structural features influencing assembly.

Main Results:

  • The miRLC is identified as the primary machinery linking pre-miRNA processing to miRNA loading.
  • The miPDC is shown to be vital for Dicer-independent miRNA biogenesis and aids in the assembly of certain Dicer-dependent miRNAs.
  • Specific pre-miRNA features (5'-uridine, 3'-mid base pairing, 5'-mid mismatches) promote miPDC assembly.

Conclusions:

  • This research provides a comprehensive understanding of miRNP/RISC assembly pathways in mammals.
  • The developed assay serves as a valuable platform for further mechanistic studies of these pathways.