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Rapid Cloning and Validation of MicroRNA Shuttle Vectors: A Practical Guide

  • Ryan L. Boudreau
  • Sara E. Garwick-Coppens
  • Jian Liu
  • Lindsay M. Wallace
  • Scott Q. HarperEmail author
Protocol
Part of the Neuromethods book series (NM, volume 58)

Abstract

MicroRNAs (miRNAs) have emerged as important modulators of eukaryotic gene expression through a process called RNA interference (RNAi). Over the last several years, a large amount of work has focused on understanding how miRNAs are expressed and processed to a biologically functional form. This knowledge has enabled the development of RNAi as a molecular tool for investigating basic biological questions or as a therapeutic technique. Artificial miRNA shuttle vectors can be engineered to mimic natural miRNAs and subsequently used to suppress any gene of interest. Here, we describe a simple method to build and functionally validate artificial miRNA shuttles.

Key words

RNAi MicroRNA miRNA, siRNA Inhibitory RNA Gene silencing Gene therapy 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ryan L. Boudreau
  • Sara E. Garwick-Coppens
  • Jian Liu
  • Lindsay M. Wallace
  • Scott Q. Harper
    • 1
    • 2
    Email author
  1. 1.Center for Gene Therapy, The Research Institute at Nationwide Children’s HospitalColumbusUSA
  2. 2.Department of Pediatrics and Molecular, Cellular, and Developmental Biology Graduate ProgramThe Ohio State University College of MedicineColumbusUSA

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