Enhancer AAVs for targeting spinal motor neurons and descending motor pathways in rodents and macaque.
Emily KussickNelson J JohansenNaz TaskinBrooke WynaldaRefugio A MartinezErin L GroceMelissa RedingElizabeth LiangLyudmila ShulgaCindy HuangTamara CasperMichael ClarkWindy HoYuan GaoCindy T J van VelthovenCassandra SobieskiRebecca FerrerMelissa R BergBritni C CurtisChris EnglishJesse C DayMichal FortunaNicholas DonadioDakota NewmanShenqin YaoAnish Bhaswanth ChakkaJeff GoldyAmy TorkelsonJunitta B GuzmanRushil ChakrabartyBeagan NguyNathan GuilfordTrangthanh H PhamVonn WrightKara RonellenfitchKathryn GudsnukBargavi ThyagarajanKimberly A SmithNick DeeHongkui ZengZizhen YaoBosiljka TasicBoaz P LeviRebecca D HodgeTrygve E BakkenEd S LeinJonathan T TingTanya L DaiglePublished in: bioRxiv : the preprint server for biology (2024)
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.
Keyphrases
- spinal cord
- single cell
- neuropathic pain
- spinal cord injury
- rna seq
- transcription factor
- endothelial cells
- binding protein
- oxidative stress
- dendritic cells
- cerebral ischemia
- regulatory t cells
- cancer therapy
- genome wide
- multiple sclerosis
- mass spectrometry
- current status
- copy number
- drug delivery
- fluorescent probe
- single molecule
- subarachnoid hemorrhage
- bone marrow