Abstract
Directed evolution represents an attractive approach to derive AAV capsid variants capable of selectively infect specific tissue or cell targets. It involves the generation of an initial library of high complexity followed by cycles of selection during which the library is progressively enriched for target-specific variants. Each selection cycle consists of the following: reconstitution of complete AAV genomes within plasmid molecules; production of virions for which each particular capsid variant is matched with the particular capsid gene encoding it; recovery of capsid gene sequences from target tissue after systemic administration. Prevalent variants are then analyzed and evaluated.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Wu Z, Asokan A, Samulski RJ (2006) Adeno-associated virus serotypes: vector toolkit for human gene therapy. Mol Ther 14:316–327
Perabo L, Endell J, King S et al (2006) Combinatorial engineering of a gene therapy vector: directed evolution of adeno-associated virus. J Gene Med 8:155–162
Maheshri N, Koerber JT, Kaspar BK et al (2006) Directed evolution of adeno-associated virus yields enhanced gene delivery vectors. Nat Biotechnol 24:198–204
Koerber JT, Maheshri N, Kaspar BK et al (2006) Construction of diverse adeno-associated viral libraries for directed evolution of enhanced gene delivery vehicles. Nat Protoc 1:701–706
Maguire CA, Gianni D, Meijer DH et al (2010) Directed evolution of adeno-associated virus for glioma cell transduction. J Neurooncol 96:337–347
Li W, Asokan A, Wu Z et al (2008) Engineering and selection of shuffled AAV genomes: a new strategy for producing targeted biological nanoparticles. Mol Ther 16:1252–1260
Grimm D, Lee JS, Wang L et al (2008) In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol 82:5887–5911
Gray SJ, Blake BL, Criswell HE et al (2010) Directed evolution of a novel adeno-associated virus (AAV) vector that crosses the seizure-compromised blood-brain barrier (BBB). Mol Ther 18:570–578
Koerber JT, Jang J-H, Schaffer DV (2008) DNA shuffling of adeno-associated virus yields functionally diverse viral progeny. Mol Ther 16:1703–1709
Müller OJ, Kaul F, Weitzman MD et al (2003) Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors. Nat Biotechnol 21:1040–1046
Michelfelder S, Lee M-K, deLima-Hahn E et al (2007) Vectors selected from adeno-associated viral display peptide libraries for leukemia cell-targeted cytotoxic gene therapy. Exp Hematol 35:1766–1776
Sellner L, Stiefelhagen M, Kleinschmidt JA et al (2008) Generation of efficient human blood progenitor-targeted recombinant adeno-associated viral vectors (AAV) by applying an AAV random peptide library on primary human hematopoietic progenitor cells. Exp Hematol 36:957–964
Naumer M, Ying Y, Michelfelder S et al (2012) Development and validation of novel AAV2 random libraries displaying peptides of diverse lengths and at diverse capsid positions. Hum Gene Ther 23:492–507
Marsic D, Govindasamy L, Currlin S et al (2014) Vector design tour de force: integrating combinatorial and rational approaches to derive novel adeno-associated virus (AAV) variants. Mol Ther 22:1900–9
Samulski RJ, Chang LS, Shenk T (1987) A recombinant plasmid from which an infectious adeno-associated virus genome can be excised in vitro and its use to study viral replication. J Virol 61:3096–3101
Gibson DG, Young L, Chuang R-Y et al (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6:343–345
Li J, Samulski RJ, Xiao X (1997) Role for highly regulated rep gene expression in adeno-associated virus vector production. J Virol 71:5236–5243
Batard P, Jordan M, Wurm F (2001) Transfer of high copy number plasmid into mammalian cells by calcium phosphate transfection. Gene 270:61–68
Vandenberghe LH, Xiao R, Lock M et al (2010) Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated virus manufacturing. Hum Gene Ther 21:1251–1257
Zolotukhin S, Byrne BJ, Mason E et al (1999) Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther 6:973–985
Arad U (1998) Modified Hirt procedure for rapid purification of extrachromosomal DNA from mammalian cells. Biotechniques 24:760–762
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Marsic, D., Zolotukhin, S. (2016). Altering Tropism of rAAV by Directed Evolution. In: Manfredsson, F. (eds) Gene Therapy for Neurological Disorders. Methods in Molecular Biology, vol 1382. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3271-9_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3271-9_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3270-2
Online ISBN: 978-1-4939-3271-9
eBook Packages: Springer Protocols