Skip to main content

Research Repository

Advanced Search

Efficient delivery of Cre-recombinase to neurons in vivo and stable transduction of neurons using adeno-associated and lentiviral vectors

Ahmed, B.Y.; Chakravarthy, S.; Eggers, R.; Hermens, W.T.; Zhang, J.Y.; Niclou, S.P.; Levelt, C.; Sablitzky, F.; Anderson, P.N.; Lieberman, A.; Verhaagen, J.

Authors

B.Y. Ahmed

S. Chakravarthy

R. Eggers

W.T. Hermens

YING ZHANG YING.ZHANG@NOTTINGHAM.AC.UK
Assistant Professor

S.P. Niclou

C. Levelt

F. Sablitzky

P.N. Anderson

A. Lieberman

J. Verhaagen



Abstract

BACKGROUND: Inactivating genes in vivo is an important technique for establishing their function in the adult nervous system. Unfortunately, conventional knockout mice may suffer from several limitations including embryonic or perinatal lethality and the compensatory regulation of other genes. One approach to producing conditional activation or inactivation of genes involves the use of Cre recombinase to remove loxP-flanked segments of DNA. We have studied the effects of delivering Cre to the hippocampus and neocortex of adult mice by injecting replication-deficient adeno-associated virus (AAV) and lentiviral (LV) vectors into discrete regions of the forebrain. RESULTS: Recombinant AAV-Cre, AAV-GFP (green fluorescent protein) and LV-Cre-EGFP (enhanced GFP) were made with the transgene controlled by the cytomegalovirus promoter. Infecting 293T cells in vitro with AAV-Cre and LV-Cre-EGFP resulted in transduction of most cells as shown by GFP fluorescence and Cre immunoreactivity. Injections of submicrolitre quantities of LV-Cre-EGFP and mixtures of AAV-Cre with AAV-GFP into the neocortex and hippocampus of adult Rosa26 reporter mice resulted in strong Cre and GFP expression in the dentate gyrus and moderate to strong labelling in specific regions of the hippocampus and in the neocortex, mainly in neurons. The pattern of expression of Cre and GFP obtained with AAV and LV vectors was very similar. X-gal staining showed that Cre-mediated recombination had occurred in neurons in the same regions of the brain, starting at 3 days post-injection. No obvious toxic effects of Cre expression were detected even after four weeks post-injection. CONCLUSION: AAV and LV vectors are capable of delivering Cre to neurons in discrete regions of the adult mouse brain and producing recombination

Citation

Ahmed, B., Chakravarthy, S., Eggers, R., Hermens, W., Zhang, J., Niclou, S., …Verhaagen, J. (2004). Efficient delivery of Cre-recombinase to neurons in vivo and stable transduction of neurons using adeno-associated and lentiviral vectors. BMC Neuroscience, 5(1), Article 4. https://doi.org/10.1186/1471-2202-5-4

Journal Article Type Article
Acceptance Date Jan 30, 2004
Online Publication Date Jan 30, 2004
Publication Date Jan 1, 2004
Deposit Date May 4, 2004
Publicly Available Date Mar 29, 2024
Journal BMC Neuroscience
Publisher Springer Verlag
Peer Reviewed Peer Reviewed
Volume 5
Issue 1
Article Number 4
DOI https://doi.org/10.1186/1471-2202-5-4
Public URL https://nottingham-repository.worktribe.com/output/1021044

Files





You might also like



Downloadable Citations