Detection of siRNA Mediated Target mRNA Cleavage Activities in Human Cells by a Novel Stem-Loop Array RT-PCR Analysis

Abstract

The small interfering RNA (siRNA)-mediated target mRNA cleavage activity generates cleaved mRNA fragments with varied termini, which creates major technical challenges for the accurate and efficient detection and verification of cleavage sites on target mRNAs. Here we used a sensitive stem-loop array reverse transcription polymerase chain reaction (SLA-RT-PCR) approach to detect and verify the siRNA-mediated target mRNA cleavage sites by determining precise sequences at the 3'- termini of cleaved mRNA fragments in human cells under physiological conditions. Our results demonstrated the great potential and broad applications of using the SLA-RT-PCR as a sensitive, cost-efficient, and high-throughput tool to systematically detect siRNA-targeted mRNA cleavage sites and fragments in human cells.

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Document Details

Document Type
Technical Report
Publication Date
Sep 07, 2016
Accession Number
AD1011348

Entities

People

  • Jack A. Roth
  • Jing Lin
  • Kai Xu
  • Lin Ji

Organizations

  • University of Texas at Austin

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Biochemistry
  • Biophysics
  • Cancer
  • Chain Reactions
  • Chemical Reactions
  • Diseases And Disorders
  • Dna Sequence Analysis
  • Dynamic Range
  • Electrophoresis
  • Gel Electrophoresis
  • Gene Expression
  • Lung Cancer
  • Molecules
  • Neoplasms
  • Pcr Testing
  • Polymerase Chain Reaction
  • Small Molecules

Fields of Study

  • Biology

Readers

  • Molecular Genetics