CoChat
Sign Up
Cited by 1,578Virus-based gene therapy researchRNA Interference and Gene DeliveryBarrier Structure and Function Studies

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

Open access
Chan, Ken Y, Jang, Min J, Yoo, Bryan B, Greenbaum, Alon, Ravi, Namita, Wu, Wei-Li, Sánchez-Guardado, Luis, Lois, Carlos +3 more·2017·Nature Neuroscience
DOI 10.1038/nn.4593PMC PMC5529245
Open this paper in CoChat — free account

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 of AAV-PHP.S transduced 82% of dorsal root ganglion neurons, as well as cardiac and enteric neurons. The efficiency of these vectors facilitates robust cotransduction 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 and enhancers, these AAVs enable efficient and targetable genetic modification of cells throughout the nervous system of transgenic and non-transgenic animals.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/nn.4593

Cites

9 of 9 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • AAVPHP.B and AAV-PHP.eB offer a more than 40-fold increase in delivery efficiency to the whole CNS.

    “For example, AAVPHP.B (146) and AAV-PHP.eB (101) offer a more than 40-fold increase in delivery efficiency to the whole CNS.”

    From Blood-Brain Barrier: From Physiology to Disease and Back (2019) · cites this paper as [101]

    The Introduction describes AAV-PHP.B as transducing neurons and astrocytes across many regions of the adult mouse brain and spinal cord, and the Results report improved CNS transduction with AAV-PHP.eB. But the Results quantify eB’s improvement over PHP.B as >2.5-fold fluorescence in the cortex and striatum and >1.8-fold in Purkinje cells; our copy does not substantiate a more-than-40-fold increase in delivery efficiency to the whole CNS for either capsid. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • AAV-PHP.B and AAV-PHP.eB show significantly higher efficiency in transporting across the BBB in C57BL/6J mice after intravenous delivery but are starkly absent in BALB/cJ mice and marmosets.

    “Gradinaru lab identified AAV-PHP⋅B and AAV-PHP.eB with significant higher efficiencies in transporting across the BBB in C57BL/6 J mice after intravenous delivery but starkly absent in BALB/cJ mice and marmosets [64–67].”

    From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [65]

    The Discussion and Results report that AAV-PHP.B was selected for BBB crossing and that intravenously delivered AAV-PHP.eB efficiently transduced mouse brain cells. Methods (Animals; page unavailable) identifies C57BL/6J mice, but the paper does not report the claimed absence of BBB transport in BALB/cJ mice or marmosets. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • AV-PHP.S is more specific for dorsal root ganglion neurons as well as cardiac and enteric neurons.

    “while AV-PHP.S is more specific for dorsal root ganglion neurons as well as and cardiac and enteric neurons (101).”

    From Blood-Brain Barrier: From Physiology to Disease and Back (2019) · cites this paper as [101]

  • Gradinaru's lab developed the CREATE platform and identified PhP.B and PhP.eB, which showed overwhelming superiority of CNS transduction after systemic delivery in some mouse strains but failed to translate into NHPs.

    “Based on such strategy, Gradinaru’s lab developed Cre recombination-based AAV targeted evolution (CREATE) platform and successfully identified PhP.B and PhP.eB [64, 65], showing overwhelming superiority of CNS transduction after systemic delivery in some mouse strains but unfortunately failed to translate into NHPs [66, 67].”

    From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [65]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 55

  1. [1]Hastie E Samulski RJ Adeno-associated virus at 50: a golden anniversary of discovery, research, and gene therapy success--a personal perspective Hum Gene Ther 26 257 265 2015 25807962 PMC4442590
  2. [2]Suzuki K In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration Nature 540 144 149 2016 27851729 PMC5331785
  3. [4]Rajasethupathy P Ferenczi E Deisseroth K Targeting Neural Circuits Cell 165 524 534 2016 27104976 PMC5296409
  4. [5]Urban DJ Roth BL DREADDs (designer receptors exclusively activated by designer drugs): chemogenetic tools with therapeutic utility Annu Rev Pharmacol Toxicol 55 399 417 2015 25292433
  5. [6]Marshall JD Cell-Type-Specific Optical Recording of Membrane Voltage Dynamics in Freely Moving Mice Cell 167 1650 1662 e1615 2016 27912066 PMC5382987
  6. [7]Kim CK Simultaneous fast measurement of circuit dynamics at multiple sites across the mammalian brain Nat Methods 13 325 328 2016 26878381 PMC5717315
  7. [8]Van der Perren A Van den Haute C Baekelandt V Viral vector-based models of Parkinson's disease Curr Top Behav Neurosci 22 271 301 2015 24839101
  8. [10]Samulski RJ Muzyczka N AAV-Mediated Gene Therapy for Research and Therapeutic Purposes Annu Rev Virol 1 427 451 2014 26958729
  9. [12]Iyer SM Virally mediated optogenetic excitation and inhibition of pain in freely moving nontransgenic mice Nat Biotechnol 32 274 278 2014 24531797 PMC3988230
  10. [13]Samad OA Virus-mediated shRNA knockdown of Na(v)1.3 in rat dorsal root ganglion attenuates nerve injury-induced neuropathic pain Mol Ther 21 49 56 2013 PMC3538302 22910296
  11. [14]Chang RB Strochlic DE Williams EK Umans BD Liberles SD Vagal Sensory Neuron Subtypes that Differentially Control Breathing Cell 161 622 633 2015 25892222 PMC4842319
  12. [16]Eldridge MA Chemogenetic disconnection of monkey orbitofrontal and rhinal cortex reversibly disrupts reward value Nat Neurosci 19 37 39 2016 26656645 PMC4866611
  13. [18]Choudhury SR Widespread Central Nervous System Gene Transfer and Silencing After Systemic Delivery of Novel AAV-AS Vector Mol Ther 24 726 735 2016 26708003 PMC4886933
  14. [19]Pulicherla N Engineering liver-detargeted AAV9 vectors for cardiac and musculoskeletal gene transfer Mol Ther 19 1070 1078 2011 21364538 PMC3129791
  15. [20]Marchio S Sidman RL Arap W Pasqualini R Brain endothelial cell-targeted gene therapy of neurovascular disorders EMBO Mol Med 8 592 594 2016 27189170 PMC4888850
  16. [21]Deverman BE Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain Nat Biotechnol 34 204 209 2016 26829320 PMC5088052
  17. [22]Betley JN Sternson SM Adeno-associated viral vectors for mapping, monitoring, and manipulating neural circuits Hum Gene Ther 22 669 677 2011 21319997 PMC3107581
  18. [23]Beier KT Circuit Architecture of VTA Dopamine Neurons Revealed by Systematic Input-Output Mapping Cell 162 622 634 2015 26232228 PMC4522312
  19. [24]Cai D Cohen KB Luo T Lichtman JW Sanes JR Improved tools for the Brainbow toolbox Nat Methods 10 540 547 2013 23866336
  20. [25]Gombash SE Intravenous AAV9 efficiently transduces myenteric neurons in neonate and juvenile mice Front Mol Neurosci 7 81 2014 25360081 PMC4197761
  21. [27]Parekh R Ascoli GA Neuronal morphology goes digital: a research hub for cellular and system neuroscience Neuron 77 1017 1038 2013 23522039 PMC3653619
  22. [28]Agha-Mohammadi S Second-generation tetracycline-regulatable promoter: repositioned tet operator elements optimize transactivator synergy while shorter minimal promoter offers tight basal leakiness J Gene Med 6 817 828 2004 15241789
  23. [29]Raymond CS Soriano P High-efficiency FLP and PhiC31 site-specific recombination in mammalian cells PLoS One 2 e162 2007 17225864 PMC1764711
  24. [30]Gallia GL Khalili K Evaluation of an autoregulatory tetracycline regulated system Oncogene 16 1879 1884 1998 9583685
  25. [32]Dimidschstein J A viral strategy for targeting and manipulating interneurons across vertebrate species Nat Neurosci 2016 PMC5348112 27798629
  26. [33]Fischer G Pan B Vilceanu D Hogan QH Yu H Sustained relief of neuropathic pain by AAV-targeted expression of CBD3 peptide in rat dorsal root ganglion Gene Ther 21 44 51 2014 24152582 PMC3881029
  27. [34]Tang JC Cell type-specific manipulation with GFP-dependent Cre recombinase Nat Neurosci 18 1334 1341 2015 26258682 PMC4839275
  28. [35]Mingozzi F High KA Immune responses to AAV vectors: overcoming barriers to successful gene therapy Blood 122 23 36 2013 23596044 PMC3701904
  29. [36]Tervo DG A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons Neuron 92 372 382 2016 27720486 PMC5872824
  30. [37]Nathanson JL Short Promoters in Viral Vectors Drive Selective Expression in Mammalian Inhibitory Neurons, but do not Restrict Activity to Specific Inhibitory Cell-Types Front Neural Circuits 3 19 2009 19949461 PMC2783723
  31. [38]Jackson KL Dayton RD Deverman BE Klein RL Better Targeting, Better Efficiency for Wide-Scale Neuronal Transduction with the Synapsin Promoter and AAV-PHP.B Front Mol Neurosci 9 116 2016 27867348 PMC5095393
  32. [39]Micheva KD Busse B Weiler NC O'Rourke N Smith SJ Single-synapse analysis of a diverse synapse population: proteomic imaging methods and markers Neuron 68 639 653 2010 21092855 PMC2995697
  33. [40]Mikuni T Nishiyama J Sun Y Kamasawa N Yasuda R High-Throughput, High-Resolution Mapping of Protein Localization in Mammalian Brain by In Vivo Genome Editing Cell 165 1803 1817 2016 27180908 PMC4912470
  34. [41]Marshel JH Mori T Nielsen KJ Callaway EM Targeting single neuronal networks for gene expression and cell labeling in vivo Neuron 67 562 574 2010 20797534 PMC2929426
  35. [42]Wertz A PRESYNAPTIC NETWORKS. Single-cell-initiated monosynaptic tracing reveals layer-specific cortical network modules Science 349 70 74 2015 26138975
  36. [44]Loulier K Multiplex cell and lineage tracking with combinatorial labels Neuron 81 505 520 2014 24507188
  37. [45]Kitamura K Judkewitz B Kano M Denk W Hausser M Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo Nat Methods 5 61 67 2008 18157136
  38. [46]Haas K Sin WC Javaherian A Li Z Cline HT Single-cell electroporation for gene transfer in vivo Neuron 29 583 591 2001 11301019
  39. [47]Young P Single-neuron labeling with inducible Cre-mediated knockout in transgenic mice Nat Neurosci 11 721 728 2008 18454144 PMC3062628
  40. [48]Badea TC New mouse lines for the analysis of neuronal morphology using CreER(T)/loxP-directed sparse labeling PLoS One 4 e7859 2009 19924248 PMC2775668
  41. [49]Lin MZ Schnitzer MJ Genetically encoded indicators of neuronal activity Nat Neurosci 19 1142 1153 2016 27571193 PMC5557009
  42. [50]Sofroniew NJ Flickinger D King J Svoboda K A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging Elife 5 2016 PMC4951199 27300105
  43. [51]Garcia AD Doan NB Imura T Bush TG Sofroniew MV GFAP-expressing progenitors are the principal source of constitutive neurogenesis in adult mouse forebrain Nat Neurosci 7 1233 1241 2004 15494728
  44. [52]Vong L Leptin action on GABAergic neurons prevents obesity and reduces inhibitory tone to POMC neurons Neuron 71 142 154 2011 21745644 PMC3134797
  45. [53]Haddad R Olfactory cortical neurons read out a relative time code in the olfactory bulb Nat Neurosci 16 949 957 2013 23685720 PMC3695490
  46. [54]Goedhart J Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93% Nat Commun 3 751 2012 22434194 PMC3316892
  47. [55]Shaner NC A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum Nat Methods 10 407 409 2013 23524392 PMC3811051
  48. [57]Oh MS Hong SJ Huh Y Kim KS Expression of transgenes in midbrain dopamine neurons using the tyrosine hydroxylase promoter Gene Ther 16 437 440 2009 18800154 PMC2747767
  49. [58]Gow A Friedrich VL Jr Lazzarini RA Myelin basic protein gene contains separate enhancers for oligodendrocyte and Schwann cell expression J Cell Biol 119 605 616 1992 1383235 PMC2289674
  50. [59]Kugler S Kilic E Bahr M Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression from an adenoviral vector in the adult rat brain depending on the transduced area Gene Ther 10 337 347 2003 12595892
  51. [60]Lee Y Messing A Su M Brenner M GFAP promoter elements required for region-specific and astrocyte-specific expression Glia 56 481 493 2008 18240313
  52. [61]Gray SJ Production of recombinant adeno-associated viral vectors and use in in vitro and in vivo administration Curr Protoc Neurosci Chapter 4, Unit 4 17 2011 21971848 PMC3209619
  53. [62]Zolotukhin S Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield Gene Ther 6 973 985 1999 10455399
  54. [63]Yang B Single-cell phenotyping within transparent intact tissue through whole-body clearing Cell 158 945 958 2014 25088144 PMC4153367
  55. [64]Hama H ScaleS: an optical clearing palette for biological imaging Nat Neurosci 18 1518 1529 2015 26368944

Cited by

Sorted by citations

Authors
Chan, K. Y., Jang, M. J., Yoo, B. B., Greenbaum, A., Ravi, N., Wu, W.-L., Sánchez-Guardado, L., Lois, C., Mazmanian, S. K., Deverman, B. E., & Gradinaru, V.
Affiliations
California Institute of Technology (US)
Corresponding author
  • Deverman, Benjamin E
  • Gradinaru, Viviana
ORCID
Venue
Nature Neuroscience
Publisher
Springer Science and Business Media LLC
Published
2017-06-26
Volume
20
Issue
8
Pages
1172-1179
ISSN
  • 1097-6256 (ISSN-L)
  • 1546-1726
DOI
10.1038/nn.4593
PMC
PMC5529245
OpenAlex
W2708579841
Cited by
1,578
References
9
Citations by year
2026: 204 · 2025: 245 · 2024: 251 · 2023: 209 · 2022: 203 · 2021: 159 · 2020: 138 · 2019: 89 · 2018: 61 · 2017: 12 · 2015: 1 · 2013: 1
FWCI
54.46
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Neuroscience (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • CaltechAUTHORS (California Institute of Technology) (submittedVersion)
Funders
  • National Institutes of Health (OT2OD023848)
  • National Institute on Aging (R01AG047664)
  • National Institute of Mental Health (R21MH103824, U01MH109147)
  • National Institute of Neurological Disorders and Stroke (DP2NS087949, U01NS090577, R01NS085910)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
Keywords
adeno-associated virus vectors, AAV capsid engineering, AAV-PHP.eB, intravenous administration, dorsal root ganglion neurons, striatal neurons, cardiac neurons, enteric neurons, cotransduction, cell type-specific promoters, transgenic animals, in vivo gene delivery
MeSH terms
Dependovirus, Animals, Ganglia, Spinal, Genetic Vectors, Mice, Transgenic, Neurons, Transduction, Genetic, Genetic Therapy, Peripheral Nervous System, Gene Transfer Techniques

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review