PUBLICATION
Highly efficient transduction of primary adult CNS and PNS neurons
- Authors
- Levin, E., Diekmann, H., Fischer, D.
- ID
- ZDB-PUB-180613-10
- Date
- 2016
- Source
- Scientific Reports 6: 38928 (Journal)
- Registered Authors
- Diekmann, Heike
- Keywords
- none
- MeSH Terms
-
- Animals
- Baculoviridae*
- Cells, Cultured
- Male
- Mice
- Mice, Transgenic
- Neurons/cytology
- Neurons/metabolism*
- Rats
- Rats, Wistar
- Transduction, Genetic/methods*
- Zebrafish
- PubMed
- 27958330 Full text @ Sci. Rep.
Citation
Levin, E., Diekmann, H., Fischer, D. (2016) Highly efficient transduction of primary adult CNS and PNS neurons. Scientific Reports. 6:38928.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping