Format

Send to

Choose Destination
Proc Natl Acad Sci U S A. 2019 Dec 9. pii: 201910061. doi: 10.1073/pnas.1910061116. [Epub ahead of print]

A systematic capsid evolution approach performed in vivo for the design of AAV vectors with tailored properties and tropism.

Author information

1
Molecular Neuromodulation, Department of Experimental Medical Science, Lund University, 221 84 Lund, Sweden.
2
Developmental and Regenerative Neurobiology, Department of Experimental Medical Science, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden.
3
Molecular Neuromodulation, Department of Experimental Medical Science, Lund University, 221 84 Lund, Sweden; tomas.bjorklund@med.lu.se.

Abstract

Adeno-associated virus (AAV) capsid modification enables the generation of recombinant vectors with tailored properties and tropism. Most approaches to date depend on random screening, enrichment, and serendipity. The approach explored here, called BRAVE (barcoded rational AAV vector evolution), enables efficient selection of engineered capsid structures on a large scale using only a single screening round in vivo. The approach stands in contrast to previous methods that require multiple generations of enrichment. With the BRAVE approach, each virus particle displays a peptide, derived from a protein, of known function on the AAV capsid surface, and a unique molecular barcode in the packaged genome. The sequencing of RNA-expressed barcodes from a single-generation in vivo screen allows the mapping of putative binding sequences from hundreds of proteins simultaneously. Using the BRAVE approach and hidden Markov model-based clustering, we present 25 synthetic capsid variants with refined properties, such as retrograde axonal transport in specific subtypes of neurons, as shown for both rodent and human dopaminergic neurons.

KEYWORDS:

barcoding; dopamine; gene therapy; retrograde transport; vector evolution

Conflict of interest statement

Competing interest statement: M.D. and T.B. are authors of a patent application covering components of the presented study.

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center