{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:14:03Z","timestamp":1785406443820,"version":"3.56.0"},"posted":{"date-parts":[[2026,1,9]]},"group-title":"Neuroscience","reference-count":34,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T00:00:00Z","timestamp":1767916800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["1RF1MH130422"],"award-info":[{"award-number":["1RF1MH130422"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["1U01AG076791"],"award-info":[{"award-number":["1U01AG076791"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Trans-neuronal viruses that spread between synaptically connected neurons have become invaluable tools in neuroscience, enabling circuit mapping and targeted delivery of genetic material to cells within defined pathways. Although most available trans-neuronal viral tracers propagate retrogradely, an anterograde trans-neuronal virus that spread from presynaptic to postsynaptic neurons would substantially expand experimental capabilities. We previously demonstrated that YFV-17D\u2014a live attenuated yellow fever vaccine used clinically for decades\u2014spreads anterogradely along neuronal circuits. We further developed a suite of recombinant YFV-17D\u2013based vectors for diverse applications, including tracing monosynaptic projectome of defined neuronal cell types, multiplex mapping of parallel pathways, and trans-neuronal genetic manipulation with minimal neurotoxicity. We have now tested and optimized procedures for vector production and application. Based on these optimizations, here, we present a two-part guide for preparing and deploying this viral vector system to map brain connectivity. The first part describes the molecular biology and cell culture workflows required to generate high-quality viral vectors, yielding titers of \u223c1 \u00d7 10^10 to 5 \u00d7 10^11 genomic copies per milliliter within approximately two weeks. The second part outlines optimized animal procedures, including intracranial injections, perioperative care, and experimental considerations tailored to specific aims and vector variants. Depending on the application, efficient trans-neuronal tracing can be achieved within 1\u20134 weeks following delivery.<\/jats:p>","DOI":"10.64898\/2026.01.08.698485","type":"posted-content","created":{"date-parts":[[2026,1,10]],"date-time":"2026-01-10T01:50:29Z","timestamp":1768009829000},"source":"Crossref","is-referenced-by-count":0,"title":["Preparation and application of YFV-17D-derived Anterograde Trans-neuronal Viral Vectors for Neuroscience Research"],"prefix":"10.64898","author":[{"given":"Tanvi","family":"Panchumarthy","sequence":"first","affiliation":[{"name":"Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lizbeth","family":"Munoz","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uzair","family":"Saleem","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ai-Vy","family":"Le","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian","family":"Nelson","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, 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