Enhancer AAVs for targeting spinal motor neurons and descending motor pathways in rodents and macaque
Brief intro:
- Author: Emily Kussick, Nelson Johansen, Naz Taskin, Brooke Wynalda, Refugio Martinez, Erin L. Groce, Melissa Reding, Elizabeth Liang, Lyudmila Shulga, Cindy Huang, Tamara Casper, Michael Clark, Windy Ho, Yuan Gao, Cindy T.J. van Velthoven, Cassandra Sobieski, Rebecca Ferrer, Melissa R. Berg, Britni C. Curtis, Chris English, Jesse C. Day, Michal Fortuna, Nicholas Donadio, Dakota Newman, Shenqin Yao, Anish Bhaswanth Chakka, Jeff Goldy, Amy Torkelson, Junitta B. Guzman, Rushil Chakrabarty, Beagen Nguy, Nathan Guilford, Trangthanh H. Pham, Vonn Wright, Kara Ronellenfitch, Kathryn Gudsnuk, Bargavi Thyagarajan, Kimberly A. Smith, Nick Dee, Hongkui Zeng, Zizhen Yao, Bosiljka Tasic, Boaz P. Levi, Rebecca Hodge, Trygve E. Bakken, Ed S. Lein, Jonathan T. Ting and Tanya L. Daigle
- Journal: BioRxiv
- Doi: https://www.doi.org/10.1101/2024.07.30.605864
- Publication Date: 2024 Jul 31
Products/Services used in the paper
Quotation shows PackGene:For injections into macaque, large scale purified viral vector preps were packaged of the PHP.eB serotype by a commercial source (Packgene, Houston TX).
Research Field:spinal motor neurons
AAV Serotype:PHP.eB
Targeted organ:spinal motor neurons
Animal or cell line strain:mice, rat, macaque
Abstract
Experimental access to cell types within the mammalian spinal cord is severely limited by the availability of genetic tools. To enable access to lower motor neurons (LMNs) and LMN subtypes, which function to integrate information from the brain and control movement through direct innervation of effector muscles, 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 (AAVs) driving a reporter fluorophore and functionally screened them in mouse. The most promising candidate enhancers were then extensively characterized using imaging and molecular techniques and further tested in rat and macaque to show conservation of LMN labeling. Additionally, we combined enhancer elements into a single vector to achieve simultaneous labeling of upper motor neurons (UMNs) and LMNs. This unprecedented LMN toolkit will enable future investigations of cell type function across species and potential therapeutic interventions for human neurodegenerative diseases.
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