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

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.

journal_name

Nat Commun

journal_title

Nature communications

authors

Adachi K,Enoki T,Kawano Y,Veraz M,Nakai H

doi

10.1038/ncomms4075

subject

Has Abstract

pub_date

2014-01-01 00:00:00

pages

3075

issn

2041-1723

pii

ncomms4075

journal_volume

5

pub_type

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