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Related Concept Videos

Alternative RNA Splicing02:18

Alternative RNA Splicing

25.4K
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
25.4K
Alternative RNA Splicing02:18

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RNA Splicing01:32

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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Related Experiment Video

Updated: Feb 25, 2026

Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
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A Cell-Based High-Throughput Method for Identifying Modulators of Alternative Splicing.

Sika Zheng1

  • 1Division of Biomedical Sciences, University of California Riverside, 201 School of Medicine Research Building, 900 University Avenue, Riverside, CA, 92521, USA. sika.zheng@ucr.edu.

Methods in Molecular Biology (Clifton, N.J.)
|August 3, 2017
PubMed
Summary
This summary is machine-generated.

Identifying regulators of alternative splicing (RAS) is crucial for understanding gene expression and disease. The novel IRAS method offers a high-throughput, sensitive, and specific approach to discover these regulators, advancing therapeutic strategies.

Keywords:
Alternative splicingDual-fluorescenceDual-outputGFPHigh-throughput screenIRASMinigeneRFPSplicing regulatorSplicing reporters

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Alternative splicing is a critical gene expression regulatory process.
  • Dysregulation of alternative splicing is linked to various human diseases.
  • Current methods for identifying splicing regulators lack throughput, sensitivity, and specificity.

Purpose of the Study:

  • To introduce IRAS (Identifying Regulators of Alternative Splicing), a novel cell-based high-throughput screening strategy.
  • To address the limitations of existing methods for discovering splicing regulators and their modulators.
  • To enable sensitive and specific identification of both positive and negative splicing regulators.

Main Methods:

  • Utilizes a pair of complementary dual-fluorescence minigene reporters (GFP and RFP) to assay distinct spliced isoforms.
  • Employs robotic liquid handlers and arrayed libraries for high-throughput screening.
  • Relies on opposite changes in GFP/RFP output ratios to confirm true splicing events and eliminate false positives.

Main Results:

  • The IRAS method achieves high throughput, high sensitivity, and high specificity in identifying splicing regulators.
  • The dual-fluorescence reporter system effectively distinguishes true splicing changes from background noise.
  • Demonstrates the capability to screen for both positive and negative splicing regulators.

Conclusions:

  • IRAS represents a significant advancement in the field of splicing regulation research.
  • This method facilitates the discovery of novel splicing regulators, paving the way for new therapeutic interventions.
  • IRAS overcomes previous limitations, offering a robust platform for exploring the complexities of alternative splicing.