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. 2025 Jun 24;44(6):115730.
doi: 10.1016/j.celrep.2025.115730. Epub 2025 May 21.

Enhancer AAVs for targeting spinal motor neurons and descending motor pathways in rodents and macaque

Emily Kussick  1 , Nelson Johansen  1 , Naz Taskin  1 , Ananya Chowdhury  1 , Meagan A Quinlan  1 , Alex Fraser  1 , Andrew G Clark  1 , Brooke Wynalda  1 , Refugio Martinez  1 , Erin L Groce  1 , Melissa Reding  1 , Elizabeth Liang  1 , Lyudmila Shulga  1 , Cindy Huang  1 , Tamara Casper  1 , Michael Clark  1 , Windy Ho  1 , Yuan Gao  1 , Cindy T J van Velthoven  1 , Cassandra Sobieski  1 , Rebecca Ferrer  1 , Melissa R Berg  2 , Britni C Curtis  2 , Chris English  2 , Jesse C Day  2 , Michal G Fortuna  1 , Nicholas Donadio  1 , Dakota Newman  1 , Shenqin Yao  1 , Anish Bhaswanth Chakka  1 , Jeff Goldy  1 , Amy Torkelson  1 , Junitta B Guzman  1 , Rushil Chakrabarty  1 , Beagen Nguy  1 , Nathan Guilford  1 , Trangthanh H Pham  1 , Vonn Wright  1 , Kara Ronellenfitch  1 , Robyn Naidoo  1 , Jaimie Kenney  1 , Ali Williford  1 , Charu Ramakrishnan  3 , Antonia Drinnenberg  3 , Kathryn Gudsnuk  1 , Bargavi Thyagarajan  1 , Kimberly A Smith  1 , Nick Dee  1 , Karl Deisseroth  4 , Hongkui Zeng  1 , Zizhen Yao  1 , Bosiljka Tasic  1 , Boaz P Levi  1 , Rebecca Hodge  1 , Trygve E Bakken  1 , Ed S Lein  1 , Jonathan T Ting  5 , Tanya L Daigle  6
Affiliations
Free article

Enhancer AAVs for targeting spinal motor neurons and descending motor pathways in rodents and macaque

Emily Kussick et al. Cell Rep. .
Free article

Abstract

Experimental access to cell types within the mammalian spinal cord is severely limited by the availability of genetic tools. To enable access to spinal motor neurons (SMNs) and SMN subtypes, we generated single-cell multiome datasets from mouse and macaque spinal cords and discovered putative enhancers for each neuronal population. We cloned these enhancers into adeno-associated viral vectors driving a reporter fluorophore and functionally screened them in the mouse. We extensively characterized the most promising candidate enhancers in rat and macaque and developed an optimized pan-SMN enhancer virus. Additionally, we generated derivative viruses expressing iCre297T recombinase or ChR2-EYFP for labeling and functional studies, and we created a single vector with combined enhancer elements to achieve simultaneous labeling of layer 5 extratelencephalic projecting neurons and SMNs. This unprecedented SMN toolkit will enable future investigations of cell type function across species and potential therapeutic interventions for human neurodegenerative diseases.

Keywords: AAV; CP: Neurosciences; cell types; enhancer; macaque; motor neuron; multiome; neurodegeneration; rodents; spinal cord.

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Conflict of interest statement

Declaration of interests T.L.D., B.P.L., B.T., E.S.L., H.Z., J.T.T., N.J., Y.G., and Z.Y. are inventors on one or more US provisional patent applications related to this work.

Update of

  • Enhancer AAVs for targeting spinal motor neurons and descending motor pathways in rodents and macaque.
    Kussick E, Johansen N, Taskin N, Chowdhury A, Quinlan MA, Fraser A, Clark AG, Wynalda B, Martinez R, Groce EL, Reding M, Liang E, Shulga L, Huang C, Casper T, Clark M, Ho W, Gao Y, van Velthoven CTJ, Sobieski C, Ferrer R, Berg MR, Curtis BC, English C, Day JC, Fortuna MG, Donadio N, Newman D, Yao S, Chakka AB, Goldy J, Torkelson A, Guzman JB, Chakrabarty R, Nguy B, Guilford N, Pham TH, Wright V, Ronellenfitch K, Naidoo R, Kenney J, Williford A, Ramakrishnan C, Drinnenberg A, Gudsnuk K, Thyagarajan B, Smith KA, Dee N, Deisseroth K, Zeng H, Yao Z, Tasic B, Levi BP, Hodge R, Bakken TE, Lein ES, Ting JT, Daigle TL. Kussick E, et al. bioRxiv [Preprint]. 2025 Mar 26:2024.07.30.605864. doi: 10.1101/2024.07.30.605864. bioRxiv. 2025. Update in: Cell Rep. 2025 Jun 24;44(6):115730. doi: 10.1016/j.celrep.2025.115730. PMID: 39131318 Free PMC article. Updated. Preprint.

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