Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain

Deverman, Benjamin E.; Pravdo, Piers L.; Simpson, Bryan P.; Kumar, Sripriya Ravindra; Chan, Ken Y.; Banerjee, Abhik; Wu, Wei; Yang, Baohua; Huber, Nina; Paşca, Sergiu P.; Gradinaru, Viviana

doi:10.1038/nbt.3440

Cited by 806 publications

(994 citation statements)

References 52 publications

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“…Cortex and CA1 typically had lower transfection efficiencies, suggesting that these regions are less susceptible to transfection after BBBO than other hippocampal fields. As a representative non-targeted region, we looked for expression in the thalamus, which was shown in previous studies to be particularly susceptible to transfection following systemic delivery of AAV9 32 , and found no significant expression (<5%, Fig. 3, a-b).…”

Section: Anatomical and Genetic Targeting Of Dreaddsmentioning

confidence: 99%

Acoustically targeted chemogenetics for the non-invasive control of neural circuits

et al. 2018

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Neurological and psychiatric diseases often involve the dysfunction of specific neural circuits in particular regions of the brain. Existing treatments, including drugs and implantable brain stimulators, aim to modulate the activity of these circuits, but are typically not cell type-specific, lack spatial targeting or require invasive procedures. Here, we introduce an approach to modulating neural circuits noninvasively with spatial, cell-type and temporal specificity. This approach, called acoustically targeted chemogenetics, or ATAC, uses transient ultrasonic opening of the blood brain barrier to transduce neurons at specific locations in the brain with virally-encoded engineered G-protein-coupled receptors, which subsequently respond to systemically administered bio-inert compounds to activate or inhibit the activity of these neurons. We demonstrate this concept in mice by using ATAC to noninvasively modify and subsequently activate or inhibit excitatory neurons within the hippocampus, showing that this enables pharmacological control of memory formation. This technology allows a brief, noninvasive procedure to make one or more specific brain regions capable of being selectively modulated using orally bioavailable compounds, thereby overcoming some of the key limitations of conventional brain therapies.

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Section: Anatomical and Genetic Targeting Of Dreaddsmentioning

confidence: 99%

Acoustically targeted chemogenetics for the non-invasive control of neural circuits

et al. 2018

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“…S7) and were electroporated using the protocol used for the cardiomyocytes once per day for 20 d. Astrocytes were fluorescently labeled with a cell-specific reporter (hGFAP::eGFP), as previously described (38). Cell morphology was followed every day during the sampling period, and despite their high overall reactivity to various stimuli and cell injuries, human astrocytes cultured on the NS tolerated the platform and the daily sampling well, with insignificant morphological changes between day 1 and day 20 (37,38).…”

Section: Pt Electrodementioning

confidence: 99%

Nondestructive nanostraw intracellular sampling for longitudinal cell monitoring

Cao

Hjort

Chen

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

129

210

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Here, we report a method for time-resolved, longitudinal extraction and quantitative measurement of intracellular proteins and mRNA from a variety of cell types. Cytosolic contents were repeatedly sampled from the same cell or population of cells for more than 5 d through a cell-culture substrate, incorporating hollow 150-nmdiameter nanostraws (NS) within a defined sampling region. Once extracted, the cellular contents were analyzed with conventional methods, including fluorescence, enzymatic assays (ELISA), and quantitative real-time PCR. This process was nondestructive with >95% cell viability after sampling, enabling long-term analysis. It is important to note that the measured quantities from the cell extract were found to constitute a statistically significant representation of the actual contents within the cells. Of 48 mRNA sequences analyzed from a population of cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs), 41 were accurately quantified. The NS platform samples from a select subpopulation of cells within a larger culture, allowing native cell-to-cell contact and communication even during vigorous activity such as cardiomyocyte beating. This platform was applied both to cell lines and to primary cells, including CHO cells, hiPSC-CMs, and human astrocytes derived in 3D cortical spheroids. By tracking the same cell or group of cells over time, this method offers an avenue to understand dynamic cell behavior, including processes such as induced pluripotency and differentiation.sampling | nanotechnology | molecular biology | cellular biology

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“…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.)…”

Section: Resultsmentioning

confidence: 99%

“…We typically use fluorescent reporters to assess gene expression in thick (≥100 µm), cleared tissue samples; below, we discuss expected results for applications presented here (Figs. 2-4) and in our previous work 2, 3 .…”

Section: Evaluation Of Transgene Expressionmentioning

confidence: 99%

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

Challis

Kumar

Chan

et al. 2018

Preprint

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We recently developed novel AAV capsids for efficient and noninvasive gene transfer across the central and peripheral nervous systems. In this protocol, we describe how to produce and systemically administer AAV-PHP viruses to label and/or genetically manipulate cells in the mouse nervous system and organs including the heart. The procedure comprises three separate stages: AAV production, intravenous delivery, and evaluation of transgene expression.The protocol spans eight days, excluding the time required to assess gene expression, and can be readily adopted by laboratories with standard molecular and cell culture capabilities. We provide guidelines for experimental design and choosing the capsid, cargo, and viral dose appropriate for the experimental aims. The procedures outlined here are adaptable to diverse biomedical applications, from anatomical and functional mapping to gene expression, silencing, and editing.

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Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain

Cited by 806 publications

References 52 publications

Acoustically targeted chemogenetics for the non-invasive control of neural circuits

Acoustically targeted chemogenetics for the non-invasive control of neural circuits

Nondestructive nanostraw intracellular sampling for longitudinal cell monitoring

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

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