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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...
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...
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...

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Related Experiment Video

Updated: May 19, 2026

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
11:00

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

Published on: June 12, 2018

Single-stranded microRNA mimics.

Guillaume Chorn1, Molly Klein-McDowell, Lihong Zhao

  • 1Sirna Therapeutics, a wholly owned subsidiary of Merck & Co., San Francisco, California 94158, USA.

RNA (New York, N.Y.)
|August 23, 2012
PubMed
Summary
This summary is machine-generated.

Single-stranded microRNAs (miRNAs) can be engineered for cellular targeting. Modifications enhance miRNA activity, paving the way for novel therapeutic mimics.

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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

Published on: September 28, 2011

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Last Updated: May 19, 2026

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

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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
07:19

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

Published on: September 28, 2011

Area of Science:

  • Molecular Biology
  • RNA Therapeutics
  • Gene Regulation

Background:

  • MicroRNAs (miRNAs) are small RNA molecules crucial for gene regulation in plants and animals.
  • miRNAs function by binding to Argonaute proteins to guide gene silencing.
  • Current miRNA therapeutics often involve double-stranded RNA, presenting delivery challenges.

Purpose of the Study:

  • To investigate the potential of single-stranded miRNAs for targeted gene regulation.
  • To identify chemical modifications and sequence features that enhance single-stranded miRNA activity.
  • To establish a foundation for developing single-stranded miRNA mimics for therapeutic applications.

Main Methods:

  • Delivery of chemically modified, 5'-phosphorylated single-stranded RNAs into cells.
  • Analysis of RNA length preferences and sensitivity to chemical modifications.
  • Genome-wide screening for seed-based targeting efficacy.
  • Optimization of passenger strand annealing with non-nucleic acid spacers.
  • Screening of random sequences to identify enhancing 3' cassette sequences.

Main Results:

  • Single-stranded miRNAs with 5'-phosphorylation and 2'-fluoro modifications demonstrated cellular targeting activity.
  • Activity was confirmed to be Argonaute (Ago)-dependent, based on length, modification sensitivity, and targeting patterns.
  • Annealing with segmented passenger strands and specific 3' pyrimidine-rich sequences significantly enhanced activity.
  • Identification of sequence elements that boost the efficacy of single-stranded miRNA mimics.

Conclusions:

  • Single-stranded miRNAs can be engineered to exhibit potent, Argonaute-mediated gene targeting.
  • Chemical modifications and specific sequence features are critical for enhancing single-stranded miRNA activity.
  • These findings represent a significant advancement in the development of single-stranded miRNA mimics for therapeutic purposes.