title: Highly efficient transduction of primary adult CNS and PNS neurons.
abstract: Delivery and expression of recombinant genes, a key methodology for many applications in biological research, remains a challenge especially for mature neurons. Here, we report easy, highly efficient and well tolerated transduction of adult peripheral and central neuronal populations of diverse species in culture using VSV-G pseudo-typed, recombinant baculovirus (BacMam). Transduction rates of up to 80% were reliably achieved at high multiplicity of infection without apparent neuro-cytopathic effects. Neurons could be transduced either shortly after plating or after several days in culture. Co-incubation with two different baculoviruses attained near complete co-localization of fluorescent protein expression, indicating multigene delivery. Finally, evidence for functional protein expression is provided by means of cre-mediated genetic recombination and neurite outgrowth assays. Recombinant protein was already detected within hours after transduction, thereby enabling functional readouts even in relatively short-lived neuronal cultures. Altogether, these results substantiate the usefulness of baculovirus-mediated transduction of mature neurons for future research in neuroscience.
authors.affiliation: Division of Experimental Neurology, Medical Faculty, Heinrich-Heine-University, Merowingerplatz 1a, 40225 Düsseldorf, Germany.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Levin
authors.affiliation: Division of Experimental Neurology, Medical Faculty, Heinrich-Heine-University, Merowingerplatz 1a, 40225 Düsseldorf, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Diekmann
authors.affiliation: Division of Experimental Neurology, Medical Faculty, Heinrich-Heine-University, Merowingerplatz 1a, 40225 Düsseldorf, Germany.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Fischer
mesh_terms: D000818:Animals; D016367:Baculoviridae; D002478:Cells, Cultured; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014161:Transduction, Genetic / Q000379:methods*; D015027:Zebrafish