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Drawing a high-resolution functional map of adeno-associated virus capsid by massively parallel sequencing

Author

Listed:
  • Kei Adachi

    (Oregon Health and Science University School of Medicine
    A portion of the study was conducted at University of Pittsburgh School of Medicine, to which K.A., T.E. and H.N. belonged until June 2011)

  • Tatsuji Enoki

    (Takara Bio Inc.
    A portion of the study was conducted at University of Pittsburgh School of Medicine, to which K.A., T.E. and H.N. belonged until June 2011)

  • Yasuhiro Kawano

    (Oregon Health and Science University School of Medicine
    Takara Bio Inc.)

  • Michael Veraz

    (Oregon Health and Science University School of Medicine)

  • Hiroyuki Nakai

    (Oregon Health and Science University School of Medicine
    A portion of the study was conducted at University of Pittsburgh School of Medicine, to which K.A., T.E. and H.N. belonged until June 2011)

Abstract

Adeno-associated virus (AAV) capsid engineering is an emerging approach to advance gene therapy. However, a systematic analysis on how each capsid amino acid contributes to multiple functions remains challenging. Here we show proof-of-principle and successful application of a novel approach, termed AAV Barcode-Seq, that allows us to characterize phenotypes of hundreds of different AAV strains in a high-throughput manner and therefore overcomes technical difficulties in the systematic analysis. In this approach, we generate DNA barcode-tagged AAV libraries and determine a spectrum of phenotypes of each AAV strain by Illumina barcode sequencing. By applying this method to AAV capsid mutant libraries tagged with DNA barcodes, we can draw a high-resolution map of AAV capsid amino acids important for the structural integrity and functions including receptor binding, tropism, neutralization and blood clearance. Thus, Barcode-Seq provides a new tool to generate a valuable resource for virus and gene therapy research.

Suggested Citation

  • Kei Adachi & Tatsuji Enoki & Yasuhiro Kawano & Michael Veraz & Hiroyuki Nakai, 2014. "Drawing a high-resolution functional map of adeno-associated virus capsid by massively parallel sequencing," Nature Communications, Nature, vol. 5(1), pages 1-14, May.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms4075
    DOI: 10.1038/ncomms4075
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