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Related Concept Videos

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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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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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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A Rapid High-throughput Method for Mapping Ribonucleoproteins RNPs on Human pre-mRNA
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Acoustic Droplet Ejection Technology and Its Application in High-Throughput RNA Interference Screening.

N Miranda Nebane1, Tatjana Coric2, Sara McKellip3

  • 1High Throughput Screening Center, Southern Research, Birmingham, AL, USA mnebane@southernresearch.org.

Journal of Laboratory Automation
|December 15, 2015
PubMed
Summary

Acoustic droplet ejection (ADE) technology enables precise, tip-free transfer of biological reagents like siRNAs in high-throughput screening (HTS). This method enhances accuracy and eliminates cross-contamination, streamlining complex screening workflows.

Keywords:
acoustic droplet ejectionsiRNA screening

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Assay Development

Background:

  • Acoustic droplet ejection (ADE) technology revolutionized liquid handling in high-throughput screening (HTS) laboratories, initially for DMSO compound solutions.
  • The Labcyte Echo 555's aqueous dispense capability expanded ADE applications to biological reagents, including siRNAs.
  • Traditional methods for siRNA screening often require step-down dilutions and are prone to liquid carryover and cross-contamination.

Purpose of the Study:

  • To describe an optimized siRNA screening platform utilizing ADE technology.
  • To demonstrate the efficacy of ADE for dispensing siRNAs in advance of assays.
  • To achieve assay parameters comparable to small-molecule screening and exceed current genomewide siRNA screen norms.

Main Methods:

  • Implementation of Labcyte's Echo 555 for precise, low-volume aqueous dispensing of siRNAs.
  • Development of a protocol for dispensing siRNA libraries weeks or months prior to assay execution.
  • Optimization of assay parameters to match small-molecule screening standards.

Main Results:

  • ADE technology, specifically the Echo 555, demonstrated accuracy and efficiency in dispensing siRNAs.
  • Elimination of liquid carryover and cross-contamination due to the tip-free nature of ADE.
  • Achieved comparable and improved assay performance metrics for siRNA screens.

Conclusions:

  • ADE technology provides a robust and efficient platform for siRNA screening.
  • The described method allows for pre-dispensing of siRNA libraries, enhancing workflow flexibility.
  • This approach significantly improves the reliability and throughput of genomewide siRNA screens.