Welcome to Viral Tool Development Core

Our organization aims to propel the study of brain fluid dynamics during sleep through the development of molecular genetics methods. We utilize genetically encoded biosensors and cell-type-specific opto-/pharmacogenetic manipulation to achieve this goal. Our core supports rodent experiments, mostly for Projects 2 and 3, by providing recombinant adeno-associated viral vectors (AAVs) tailored to label key fluid compartments like interstitial fluid, cerebrospinal fluid (CSF), and blood. Our approach enables longitudinal imaging in existing transgenic mice. Additionally, we design transgenic mouse constructs for visualizing vascular and perivascular dynamics.

Research Aims

Aim 1: Establish a viral vector pipeline for imagaing and manipulation 

Aim 2: Develop genetically encoded fluorescent plasma labels

Aim 3: Extend the utility of plasma label with biosensor attachment 

Aim 4: Stable expression of fluid probes by one-shot gene editing

Publications

Asiminas A, Gomolka RS, Gregoriades S, Hirase H, Nedergaard M, Beinlich FRM. Protocol to study oxygen dynamics in the in vivo mouse brain using bioluminescence microscopy.  STAR Protoc. 2024 Sep 25;5(4):103334. doi: 10.1016/j.xpro.2024.103334. Online ahead of print. PMID: 39331498 Free PMC article.

Vittani M, Knak PAG, Fukuda M, Nagao M, Wang X, Kjaerby C, Konno A, Hirai H, Nedergaard M, Hirase H. Virally induced CRISPR/Cas9-based knock-in of fluorescent albumin allows long-term visualization of cerebral circulation in infant and adult mice. Neuromethods, 2025; 209:127-144. doi: 10.1007/978-1-0716-4011-1_6

Beinlich FRM, Asiminas A, Untiet V, Bojarowska Z, Plá V, Sigurdsson B, Timmel V, Gehrig L, Graber MH, Hirase H, Nedergaard M. Oxygen imaging of hypoxic pockets in the mouse cerebral cortex. Science. 2024 Mar 29;383(6690):1471-1478. doi: 10.1126/science.adn1011. Epub 2024 Mar 28. PMID: 38547288 Free PMC article.

Hussain R, Tithof J, Wang W, Cheetham-West A, Song W, Peng W, Sigurdsson B, Kim D, Sun Q, Peng S, Plá V, Kelley DH, Hirase H, Castorena-Gonzalez JA, Weikop P, Goldman SA, Davis MJ, Nedergaard M. Potentiating glymphatic drainage minimizes post-traumatic cerebral oedema. Nature. 2023 Nov;623(7989):992-1000. doi: 10.1038/s41586-023-06737-7. Epub 2023 Nov 15. PMID: 37968397 Free PMC article.

Nasu Y, Aggarwal A, Le GNT, Vo CT, Kambe Y, Wang X, Beinlich FRM, Lee AB, Ram TR, Wang F, Gorzo KA, Kamijo Y, Boisvert M, Nishinami S, Kawamura G, Ozawa T, Toda H, Gordon GR, Ge S, Hirase H, Nedergaard M, Paquet ME, Drobizhev M, Podgorski K, Campbell RE. Lactate biosensors for spectrally and spatially multiplexed fluorescence imaging. Nat Commun. 2023 Oct 27;14(1):6598. doi: 10.1038/s41467-023-42230-5. PMID: 37891202 Free PMC article.

Dietz AG, Weikop P, Hauglund N, Andersen M, Petersen NC, Rose L, Hirase H, Nedergaard M. Local extracellular K+ in cortex regulates norepinephrine levels, network state, and behavioral output. Proc Natl Acad Sci U S A. 2023 Oct 3;120(40):e2305071120. doi: 10.1073/pnas.2305071120. Epub 2023 Sep 29. PMID: 37774097 Free PMC article.