RNA trans-splicing for genodermatoses

Methods Mol Biol. 2013:961:441-55. doi: 10.1007/978-1-62703-227-8_30.

Abstract

Spliceosome-mediated RNA trans-splicing (SMaRT) is a tool that facilitates the recombination of two distinct pre-mRNA molecules. Its application for gene therapeutic purposes has been hindered by laborious procedures to identify gene-specific molecules. We have established a screening method for the identification of highly functional RNA trans-splicing molecules based on fluorescence reporters, facilitating the generation of most potent therapeutic molecules for the correction of any gene of interest.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Culture Techniques / methods
  • Cell Line
  • Cloning, Molecular / methods
  • DNA Primers / genetics
  • Flow Cytometry / methods
  • Genes, Reporter
  • Genetic Therapy / methods*
  • Green Fluorescent Proteins / genetics
  • Humans
  • Luminescent Proteins / genetics
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods
  • RNA, Messenger / genetics
  • Skin / metabolism
  • Skin Diseases / genetics*
  • Skin Diseases / therapy
  • Spliceosomes / genetics
  • Trans-Splicing*
  • Transfection

Substances

  • DNA Primers
  • Luminescent Proteins
  • RNA, Messenger
  • fluorescent protein 583
  • Green Fluorescent Proteins