Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems

Abstract

Adeno-associated viruses (AAVs) are commonly used for in vivo gene transfer. Nevertheless, AAVs that provide efficient transduction across specific organs or cell populations are needed. Here, we describe AAV-PHP.eB and AAV-PHP.S, capsids that efficiently transduce the central and peripheral nervous systems, respectively. In the adult mouse, intravenous administration of 1×10 11 vector genomes (vg) of AAV-PHP.eB transduced 69% of cortical and 55% of striatal neurons, while 1×10 12 vg AAV-PHP.S transduced 82% of dorsal root ganglion neurons, as well as cardiac and enteric neurons. The efficiency of these vectors facilitates robust co-transduction and stochastic, multicolor labeling for individual cell morphology studies. To support such efforts, we provide methods for labeling a tunable fraction of cells without compromising color diversity. Furthermore, when used with cell type-specific promoters, these AAVs provide targeted gene expression across the nervous system and enable efficient and versatile gene manipulation throughout the nervous system of transgenic and non-transgenic animals.Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

“…Use sequence editing and annotation software to determine the unique attributes of the rAAV genome. In Table 1, we list genomes used here and in our previous work 2,3 ; see also Addgene's plasmid repository for pAAVs that may be The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/246405 doi: bioRxiv preprint first posted online Jan. 11, 2018; (Sup.)…”

“…Fluorescent reporters are commonly used for cell type-specific mapping and phenotyping 2,38,39 (Figs. 2-4).…”

“…Use sequence editing and annotation software to determine the unique attributes of the rAAV genome. In Table 1, we list genomes used here and in our previous work 2,3 ; see also Addgene's plasmid repository for pAAVs that may be suitable for different applications. DREADDs = Designer Receptors Exclusively Activated by Designed Drugs; CRISPR = Clustered Regularly Interspaced Short Palindromic Repeats; shRNA = short hairpin RNA.…”

“…AAV-PHP.eB and AAV-PHP.S produce viral yields similar to other high producing naturally occurring serotypes (e.g., AAV9) and enable efficient, noninvasive gene transfer to the CNS or PNS and visceral organs, respectively 2 …”

Circumventing the blood–brain barrier

“…At the California Institute of Technology in Pasadena, Viviana Gradinaru's group developed a high-throughput system for generating mutations in the shell of AAV9 (which helps the virus target different tissues and cells) and testing millions of these variations (6). In June, her team discovered one highly efficient version that, at relatively low intravenous doses, can deliver genes to about 70% of neurons in the cerebral cortex of adult mice (7). Gradinaru plans to use the virus as a basic research tool but says the finding could lay the groundwork for potential gene therapies in the future.…”