| First Author | Levin E | Year | 2016 |
| Journal | Sci Rep | Volume | 6 |
| Pages | 38928 | PubMed ID | 27958330 |
| Mgi Jnum | J:254376 | Mgi Id | MGI:6101542 |
| Doi | 10.1038/srep38928 | Citation | Levin E, et al. (2016) Highly efficient transduction of primary adult CNS and PNS neurons. Sci Rep 6:38928 |
| abstractText | 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. |
