Lentiviral vector-mediated delivery of short hairpin RNA results in persistent knockdown of gene expression in mouse brain.
RNA interference (RNAi) is an evolutionarily conserved mechanism of posttranscriptional gene-specific silencing. For in vivo applications, RNAi has been hampered until recently by inefficient delivery methods and by the transient nature of the gene suppression. Lentiviral vectors (LVs) hold great promise for gene therapeutic applications, pharmaceutical target validation, and functional genomics because stable gene transfer is mediated both in dividing and nondividing cells. We have used a lentiviral vector-based system for RNAi. We produced human immunodeficiency virus type 1-derived LVs encoding a short hairpin RNA specific for enhanced green fluorescent protein (EGFP) mRNA that were capable of inhibiting EGFP expression in mammalian cells. EGFP knockdown persisted after multiple passages of the cells. Of particular interest, our RNAi LVs were equally effective in suppression and prevention of EGFP expression after stereotactic injection in adult mouse brain. Therefore, we believe that the use of LVs for stable RNAi in brain will become a powerful aid to probe gene function in vivo and for gene therapy of diseases of the central nervous system.
Van den Haute
Chris
C
Gene Therapy Program, Experimental Neurosurgery and Neuroanatomy, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium.
Eggermont
Kristel
K
Nuttin
Bart
B
Debyser
Zeger
Z
Baekelandt
Veerle
V
D000818:Animals; D001483:Base Sequence; D001921:Brain; D016543:Central Nervous System Neoplasms / Q000235:genetics / Q000628:therapy; D015536:Down-Regulation; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins