TY - JOUR
T1 - A versatile viral toolkit for functional discovery in the nervous system
AU - Pouchelon, Gabrielle
AU - Vergara, Josselyn
AU - McMahon, Justin
AU - Gorissen, Bram L.
AU - Lin, Jessica D.
AU - Vormstein-Schneider, Douglas
AU - Niehaus, Jason L.
AU - Burbridge, Timothy J.
AU - Wester, Jason C.
AU - Sherer, Mia
AU - Fernandez-Otero, Marian
AU - Allaway, Kathryn C.
AU - Pelkey, Kenneth
AU - Chittajallu, Ramesh
AU - McBain, Chris J.
AU - Fan, Melina
AU - Nasse, Jason S.
AU - Wildenberg, Gregg A.
AU - Fishell, Gordon
AU - Dimidschstein, Jordane
N1 - Funding Information:
We thank S. Gerard, P. Delvenne, C. Parotte, D. Joseph, O. Lis, P. Copine, Ma. Matou, P. Noir, Mi. Mi, and Co. Co. for their useful comments on the manuscript. We also thank Kimberly Ritola for sharing EnvA-N2c-dG rabies and her advice on recombination-free AAV production and Bernardo Rudy, Robert Machold, and Benjamin Schuman for discussing their VTKS5 experiment using NPY-FlpO animals. J.D. is supported by NIH grants R01-MH111529 and UG3MH120096 in addition to support from Simons Foundation Award 566615. G.P. was supported by an EMBO Long-Term fellowship, early and advanced Swiss Foundation postdoctoral fellowships, and a Hearst foundation grant, and G.F. was supported by grants from the NIH (MH071679, NS08297, NS074972, and MH095147) as well as support from the Simons Foundation (SFARI). G.P. and J.D. conceived the project. G.P. J.D. and G.F. wrote the manuscript. G.P. J.V. and J.M. performed injections and the neuron-specificity experiments. G.P. performed the tracing and c-Fos experiments. J.D.L. and D.V.-S. performed the interference experiments. T.J.B. performed calcium imaging. G.A.W. performed electron microscopy. K.C.A. M.F.-O. and M.S. performed the CRISPR-based experiment. K.P. R.C. J.C.W. and C.J.M. injected mice and performed optogenetic recordings. G.P. J.V. J.M. and B.L.G. analyzed the Addgene data and prepared the figures. M.F. J.L.N. and J.S.N. provided the sequence data for all Addgene plasmids used in this study. J.D. is founder, shareholder, and employee of Regel Therapeutics. G.F. is founder and shareholder of Regel Therapeutics. The work presented in this manuscript is unrelated to any aspect of Regel Therapeutics’ focus, work, or interest. We worked to ensure sex balance in the selection of non-human subjects. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in science. One or more of the authors of this paper self-identifies as a member of the LGBTQ+ community.
Funding Information:
We thank S. Gerard, P. Delvenne, C. Parotte, D. Joseph, O. Lis, P. Copine, Ma. Matou, P. Noir, Mi. Mi, and Co. Co. for their useful comments on the manuscript. We also thank Kimberly Ritola for sharing EnvA-N2c-dG rabies and her advice on recombination-free AAV production and Bernardo Rudy, Robert Machold, and Benjamin Schuman for discussing their VTKS5 experiment using NPY-FlpO animals. J.D. is supported by NIH grants R01-MH111529 and UG3MH120096 in addition to support from Simons Foundation Award 566615 . G.P. was supported by an EMBO Long-Term fellowship, early and advanced Swiss Foundation postdoctoral fellowships, and a Hearst foundation grant, and G.F. was supported by grants from the NIH ( MH071679 , NS08297 , NS074972 , and MH095147 ) as well as support from the Simons Foundation (SFARI).
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/6/20
Y1 - 2022/6/20
N2 - The ability to precisely control transgene expression is essential for basic research and clinical applications. Adeno-associated viruses (AAVs) are non-pathogenic and can be used to drive stable expression in virtually any tissue, cell type, or species, but their limited genomic payload results in a trade-off between the transgenes that can be incorporated and the complexity of the regulatory elements controlling their expression. Resolving these competing imperatives in complex experiments inevitably results in compromises. Here, we assemble an optimized viral toolkit (VTK) that addresses these limitations and allows for efficient combinatorial targeting of cell types. Moreover, their modular design explicitly enables further refinements. We achieve this in compact vectors by integrating structural improvements of AAV vectors with innovative molecular tools. We illustrate the potential of this approach through a systematic demonstration of their utility for targeting cell types and querying their biology using a wide array of genetically encoded tools.
AB - The ability to precisely control transgene expression is essential for basic research and clinical applications. Adeno-associated viruses (AAVs) are non-pathogenic and can be used to drive stable expression in virtually any tissue, cell type, or species, but their limited genomic payload results in a trade-off between the transgenes that can be incorporated and the complexity of the regulatory elements controlling their expression. Resolving these competing imperatives in complex experiments inevitably results in compromises. Here, we assemble an optimized viral toolkit (VTK) that addresses these limitations and allows for efficient combinatorial targeting of cell types. Moreover, their modular design explicitly enables further refinements. We achieve this in compact vectors by integrating structural improvements of AAV vectors with innovative molecular tools. We illustrate the potential of this approach through a systematic demonstration of their utility for targeting cell types and querying their biology using a wide array of genetically encoded tools.
KW - AAV
KW - circuits
KW - neuroscience
KW - vector
UR - http://www.scopus.com/inward/record.url?scp=85132744883&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85132744883&partnerID=8YFLogxK
U2 - 10.1016/j.crmeth.2022.100225
DO - 10.1016/j.crmeth.2022.100225
M3 - Article
AN - SCOPUS:85132744883
SN - 2667-2375
VL - 2
JO - Cell Reports Methods
JF - Cell Reports Methods
IS - 6
M1 - 100225
ER -