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Chimeric RNA pp 219-232 | Cite as

SMaRT for Therapeutic Purposes

  • Lisa M. RiedmayrEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2079)

Abstract

Spliceosome-mediated mRNA trans-splicing (SMaRT) is a promising strategy for treatment of genetic diseases which cannot be targeted via classical therapy approaches. SMaRT utilizes an exogenous pre-mRNA trans-splicing molecule (PTM) to correct a diseased target pre-mRNA. This process relies on splicing of two separate pre-mRNA molecules in trans creating a mature chimeric mRNA molecule which consists of the protein coding sequence of the PTM as well as the endogenous mRNA. For therapeutic implications, the most critical step in SMaRT is to develop PTMs resulting in a high ratio of trans-splicing to regular cis-splicing.

This protocol provides guidelines on how to design PTMs and describes a fast screening assay to test their efficiencies. To elucidate the therapeutic potential of the best candidates in a more native setting, these PTMs are tested further on mini genes.

Key words

SMaRT Trans-splicing Gene therapy AAV Gene correction Chimeric mRNA 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Center for Integrated Protein Science Munich CIPSMLudwig-Maximilians-Universität MünchenMunichGermany
  2. 2.Department of Pharmacy, Center for Drug ResearchLudwig-Maximilians-Universität MünchenMunichGermany

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