Robust in vivo gene transfer into adult mammalian neural stem cells by lentiviral vectors.
Stable genetic modification of adult stem cells is fundamental for both developmental studies and therapeutic purposes. Using in vivo marking studies, we showed that injection of lentiviral vectors (LVs) into the subventricular zone of the adult mouse brain enables efficient gene transfer into long-term self-renewing neural precursors and steady, robust vector expression in their neuronal progeny throughout the subventricular zone and its rostral extension, up to the olfactory bulb. By clonal and population analysis in culture, we proved that in vivo-marked neural precursors display self-renewal and multipotency, two essential characteristics of neural stem cells (NSCs). Thus, LVs efficiently target long-term repopulating adult NSCs, and the effect of the initial transduction is amplified by the continuous generation of NSC-derived, transduced progeny. LVs may thus allow novel studies on NSCs' physiology in vivo, and introduction of therapeutic genes into NSCs may allow the development of novel approaches for untreatable CNS diseases.
Consiglio
Antonella
A
San Raffaele Telethon Institute for Gene Therapy, Milan, Italy.
Gritti
Angela
A
Dolcetta
Diego
D
Follenzi
Antonia
A
Bordignon
Claudio
C
Gage
Fred H
FH
Vescovi
Angelo Luigi
AL
Naldini
Luigi
L
D000818:Animals; D001921:Brain / Q000166:cytology / Q000502:physiology; D018929:Cell Culture Techniques; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D011994:Recombinant Proteins / Q000032:analysis; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014162:Transfection / Q000379:methods
D011994:Recombinant Proteins