Directed Evolution of a Novel Adeno-associated Virus (AAV) Vector That Crosses the Seizure-compromised Blood–Brain Barrier (BBB)

DNA shuffling and directed evolution were employed to develop a novel adeno-associated virus (AAV) vector capable of crossing the seizure-compromised blood–brain barrier (BBB) and transducing cells in the brain. Capsid DNA from AAV serotypes 1–6, 8, and 9 were shuffled and recombined to create a lib...

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Published in:Molecular therapy 2010-03, Vol.18 (3), p.570-578
Main Authors: Gray, Steven J, Blake, Bonita L, Criswell, Hugh E, Nicolson, Sarah C, Samulski, R Jude, McCown, Thomas J
Format: Article
Language:English
Subjects:
Amino acids
Animals
Blood-Brain Barrier - chemistry
Brain
Capsid - chemistry
Cell Line
Cell Survival
Cloning
Convulsions & seizures
Dependovirus - metabolism
Directed Molecular Evolution
DNA - metabolism
Epilepsy
Female
Gene therapy
Genetic Therapy - methods
Green Fluorescent Proteins - chemistry
Humans
Immunohistochemistry - methods
Kainic Acid - chemistry
Library management
Mice
Mice, Inbred BALB C
Microscopy, Confocal - methods
Mutation
Neurons - metabolism
Original
Rats
Seizures - metabolism
Vectors (Biology)
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cited_by cdi_FETCH-LOGICAL-c478t-84aac28da8cb5c3e048099516dfbbb30d852eea05548564df6183afa1406a2013
cites cdi_FETCH-LOGICAL-c478t-84aac28da8cb5c3e048099516dfbbb30d852eea05548564df6183afa1406a2013
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container_issue 3
container_start_page 570
container_title Molecular therapy
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creator Gray, Steven J
Blake, Bonita L
Criswell, Hugh E
Nicolson, Sarah C
Samulski, R Jude
McCown, Thomas J
description DNA shuffling and directed evolution were employed to develop a novel adeno-associated virus (AAV) vector capable of crossing the seizure-compromised blood–brain barrier (BBB) and transducing cells in the brain. Capsid DNA from AAV serotypes 1–6, 8, and 9 were shuffled and recombined to create a library of chimeric AAVs. One day after kainic acid–induced limbic seizure activity in rats, the virus library was infused intravenously (i.v.), and 3 days later, neuron-rich cells were mechanically dissociated from seizure-sensitive brain sites, collected and viral DNA extracted. After three cycles of selection, green fluorescent protein (GFP)–packaged clones were administered directly into brain or i.v. 1 day after kainic acid–induced seizures. Several clones that were effective after intracranial administration did not transduce brain cells after the i.v. administration. However, two clones (32 and 83) transduced the cells after direct brain infusion and after i.v. administration transduced the cells that were localized to the piriform cortex and ventral hippocampus, areas exhibiting a seizure-compromised BBB. No transduction occurred in areas devoid of BBB compromise. Only one parental serotype (AAV8) exhibited a similar expression profile, but the biodistribution of 32 and 83 diverged dramatically from this parental serotype. Thus, novel AAV vectors have been created that can selectively cross the seizure-compromised BBB and transduce cells.
doi_str_mv 10.1038/mt.2009.292
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source PubMed (Medline); ScienceDirect®
subjects Amino acids
Animals
Blood-Brain Barrier - chemistry
Brain
Capsid - chemistry
Cell Line
Cell Survival
Cloning
Convulsions & seizures
Dependovirus - metabolism
Directed Molecular Evolution
DNA - metabolism
Epilepsy
Female
Gene therapy
Genetic Therapy - methods
Green Fluorescent Proteins - chemistry
Humans
Immunohistochemistry - methods
Kainic Acid - chemistry
Library management
Mice
Mice, Inbred BALB C
Microscopy, Confocal - methods
Mutation
Neurons - metabolism
Original
Rats
Seizures - metabolism
Vectors (Biology)
title Directed Evolution of a Novel Adeno-associated Virus (AAV) Vector That Crosses the Seizure-compromised Blood–Brain Barrier (BBB)
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