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      Brain-wide TVA compensation allows rabies virus to retrograde target cell-type-specific projection neurons

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          Abstract

          Retrograde tracers based on viral vectors are powerful tools for the imaging and manipulation of upstream neural networks projecting to a specific brain region, and they play important roles in structural and functional studies of neural circuits. However, currently reported retrograde viral tracers have many limitations, such as brain area selectivity or the inability to retrograde label genetically defined brain-wide projection neurons. To overcome these limitations, a new retrograde tracing method, AAV-PHP.eB assisted retrograde tracing systems (PARTS) based on rabies virus, was established through brain-wide TVA-dependent targeting using an AAV-PHP.eB that efficiently crosses the blood–brain barrier in C57BL/6 J mice, and complementation of EnvA-pseudotyped defective rabies virus that specifically recognizes the TVA receptor. Furthermore, combined with Cre transgenic mice, cell-type-specific PARTS (cPARTS) was developed, which can retrograde label genetically defined brain-wide projection neurons. Our research provides new tools and technical support for the analysis of neural circuits.

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

          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 1×1011 vector genomes (vg) of AAV-PHP.eB transduced 69% of cortical and 55% of striatal neurons, while 1×1012 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.
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            A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons

            Efficient retrograde access to projection neurons for the delivery of sensors and effectors constitutes an important and enabling capability for neural circuit dissection. Such an approach would also be useful for gene therapy, including the treatment of neurodegenerative disorders characterized by pathological spread through functionally connected and highly distributed networks. Viral vectors, in particular, are powerful gene delivery vehicles for the nervous system, but all available tools suffer from inefficient retrograde transport or limited clinical potential. To address this need, we applied in vivo directed evolution to engineer potent retrograde functionality into the capsid of adeno-associated virus (AAV), a vector that has shown promise in neuroscience research and the clinic. A newly evolved variant, rAAV2-retro, permits robust retrograde access to projection neurons with efficiency comparable to classical synthetic retrograde tracers and enables sufficient sensor/effector expression for functional circuit interrogation and in vivo genome editing in targeted neuronal populations. VIDEO ABSTRACT.
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              Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain

              Recombinant adeno-associated viruses (rAAVs) are commonly used vehicles for in vivo gene transfer 1-6 . However, the tropism repertoire of naturally occurring AAVs is limited, prompting a search for novel AAV capsids with desired characteristics 7-13 . Here we describe a capsid selection method, called Cre-recombination-based AAV targeted evolution (CREATE), that enables the development of AAV capsids that more efficiently transduce defined Cre-expressing cell populations in vivo. We use CREATE to generate AAV variants that efficiently and widely transduce the adult mouse central nervous system (CNS) after intravenous injection. One variant, AAV-PHP.B, transfers genes throughout the CNS with an efficiency that is at least 40-fold greater than that of the current standard, AAV9 14-17 , and transduces the majority of astrocytes and neurons across multiple CNS regions. In vitro, it transduces human neurons and astrocytes more efficiently than does AAV9, demonstrating the potential of CREATE to produce customized AAV vectors for biomedical applications.
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                Author and article information

                Contributors
                kz.lin@siat.ac.cn
                fq.xu@siat.ac.cn
                Journal
                Mol Brain
                Mol Brain
                Molecular Brain
                BioMed Central (London )
                1756-6606
                29 January 2022
                29 January 2022
                2022
                : 15
                : 13
                Affiliations
                [1 ]GRID grid.458518.5, ISNI 0000 0004 1803 4970, Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, , National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences-Wuhan National Laboratory for Optoelectronics, ; Wuhan, 430071 People’s Republic of China
                [2 ]GRID grid.410726.6, ISNI 0000 0004 1797 8419, University of Chinese Academy of Sciences, ; Beijing, 100049 People’s Republic of China
                [3 ]GRID grid.9227.e, ISNI 0000000119573309, The Brain Cognition and Brain Disease Institute (BCBDI), Shenzhen Key Laboratory of Viral Vectors for Biomedicine, Shenzhen Institute of Advanced Technology, , Chinese Academy of Sciences, ; Shenzhen, 518055 People’s Republic of China
                [4 ]GRID grid.458489.c, ISNI 0000 0001 0483 7922, Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions, NMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal Products, Shenzhen, Key Laboratory of Quality Control Technology for Virus-Based Therapeutics, Guangdong Provincial Medical Products Administration, ; Shenzhen, 518055 People’s Republic of China
                [5 ]GRID grid.33199.31, ISNI 0000 0004 0368 7223, Wuhan National Laboratory for Optoelectronics, , Huazhong University of Science and Technology, ; Wuhan, 430074 People’s Republic of China
                [6 ]GRID grid.33199.31, ISNI 0000 0004 0368 7223, Department of Pathophysiology, Key Lab of Neurological Disorder of Education Ministry, School of Basic Medicine, , Tongji Medical College, Huazhong University of Science and Technology, ; Wuhan, People’s Republic of China
                [7 ]GRID grid.49470.3e, ISNI 0000 0001 2331 6153, College of Life Sciences, , Wuhan University, ; Wuhan, People’s Republic of China
                [8 ]GRID grid.49470.3e, ISNI 0000 0001 2331 6153, HongYi Honor College, Wuhan University, ; Wuhan, People’s Republic of China
                [9 ]GRID grid.458489.c, ISNI 0000 0001 0483 7922, Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions, ; Shenzhen, 518055 People’s Republic of China
                [10 ]GRID grid.9227.e, ISNI 0000000119573309, Center for Excellence in Brain Science and Intelligence Technology, , Chinese Academy of Sciences, ; Shanghai, 200031 People’s Republic of China
                Author information
                http://orcid.org/0000-0001-5091-6197
                Article
                898
                10.1186/s13041-022-00898-8
                8800268
                35093138
                6e52ea80-bbfd-438e-8e98-1a96ac6d4d80
                © The Author(s) 2022

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                : 26 October 2021
                : 16 January 2022
                Funding
                Funded by: Key-Area Research and Development Program of Guangdong Province
                Award ID: 2018B030331001
                Award Recipient :
                Funded by: Strategic Priority Research Program of the Chinese Academy of Sciences
                Award ID: XDB32030200
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31830035
                Award ID: 31771156
                Award ID: 21921004
                Award Recipient :
                Funded by: Shenzhen Key Laboratory of Viral Vectors for Biomedicine
                Award ID: ZDSYS20200811142401005
                Award Recipient :
                Funded by: National Key R&D Program of China
                Award ID: 2018YFC1704600
                Award Recipient :
                Categories
                Methodology
                Custom metadata
                © The Author(s) 2022

                Neurosciences
                retrograde tracing,rabies virus,aav-php.eb,cell-type-specific neurons,projection neurons

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