Adeno-associated virus effectively mediates conditional gene modification in the brain.
The Cre/loxP system is increasingly showing promise for investigating genes involved in neural function. Here, we demonstrate that in vivo modification of genes in the mouse brain can be accomplished in a spatial- and temporal-specific manner by targeted delivery of an adeno-associated virus (AAV) encoding a green fluorescent protein/Cre recombinase (GFP/Cre) fusion protein. By using a reporter mouse, in which Cre recombinase activates beta-galactosidase expression, we demonstrate long-term recombination of neurons in the hippocampus, striatum, and septum as early as 7 days after stereotaxic injection of virus. Recombined cells were observed for at least 6 months postinjection without evidence of cell loss or neural damage. AAV-mediated delivery of GFP/Cre provides a valuable approach to alter the mouse genome, as AAV delivers genes efficiently to neurons with low toxicity. This approach will greatly facilitate the study of genetic modifications in the mouse brain.
Kaspar
Brian K
BK
Laboratory of Genetics, Molecular Neurobiology Laboratory, and Peptide Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Vissel
Bryce
B
Bengoechea
Tasha
T
Crone
Steven
S
Randolph-Moore
Lynne
L
Muller
Rolf
R
Brandon
Eugene P
EP
Schaffer
David
D
Verma
Inder M
IM
Lee
Kuo-Fen
KF
Heinemann
Stephen F
SF
Gage
Fred H
FH
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D003342:Corpus Striatum / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000378:metabolism; D006720:Homozygote; D006801:Humans; D007150:Immunohistochemistry; D019426:Integrases / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D020665:Septum of Brain / Q000378:metabolism; D013997:Time Factors; D014764:Viral Proteins / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase