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Design of AAV Vectors for Delivery of RNAi

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Adeno-Associated Virus Vectors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1950))

Abstract

Adeno-associated viral vectors have emerged as an important tool for human gene therapy, having demonstrated high transduction efficiency in a broad range of target tissues, a good safety profile in animal models and human clinical trials, and prospective long-lasting gene expression. First discovered 20 years ago, RNA interference (RNAi) has become another important tool for human gene therapy, enabling scientists to move on from classical gene transfer to gene silencing approaches, or combinations thereof. In this chapter, we describe a simple step-by-step method that will allow gene silencing novices to design their own artificial miRNAs against a target of their choice, clone these miRNAs into an AAV-based vector, and rapidly screen for highly efficient artificial miRNAs. The described method takes into consideration recent advances in the field including miRNA processing from various cellular miRNA backbones, choice between polymerase II and III promoters, and the potential impact of these factors on toxicity as it relates to off-targeting and to saturation of the endogenous RNAi machinery.

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Author information

Authors and Affiliations

  1. Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA

    Florie Borel & Christian Mueller

  2. Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA, USA

    Christian Mueller

Authors
  1. Florie Borel
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  2. Christian Mueller
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Corresponding author

Correspondence to Christian Mueller .

Editor information

Editors and Affiliations

  1. Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA

    Michael J. Castle

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Borel, F., Mueller, C. (2019). Design of AAV Vectors for Delivery of RNAi. In: Castle, M. (eds) Adeno-Associated Virus Vectors. Methods in Molecular Biology, vol 1950. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9139-6_1

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  • DOI: https://doi.org/10.1007/978-1-4939-9139-6_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9138-9

  • Online ISBN: 978-1-4939-9139-6

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