Biology Reference
In-Depth Information
Chapter 1
Lentiviral Vectors as Research Tools in Neurobiology:
Design and Production
Alessandro Papale and Riccardo Brambilla
Abstract
Viral vectors are now common in contemporary neuroscience research and their use as gene transfer tools
for the central nervous system has seen an enormous growth in the last 2 decades. This chapter discusses
about designing, production, and use of lentiviral vectors (LVs), one of the most popular and versatile
system currently available.
Key words Viral vector, Neuroscience, Gene transfer, Gene silencing, Inducible vectors
1
Introduction
Replication-defective lentiviral vectors (LVs) were originally
derived from HIV-1, a virus of the Lentivirus genus. Lentiviruses
are complex viruses characterized by a unique virion morphology,
with cylindrical or conical cores [ 1 ]. The wild-type virus is an
enveloped single-stranded RNA virus with approximately 9 kb
of genome. During the infection cycle, his RNA-based genome is
converted in dsDNA and integrates in the host; this characteristic
is conserved in the vector and allows very long-term expression of
the transgene. Wild-type HIV-1 is the pathogenic agent of
Acquired Immune Deficiency Syndrome, and the first developed
LVs were originally developed to transduce macrophages, lympho-
cytes, and dendritic cells, to track viral replication and as platforms
to screen for anti-HIV-1 drugs [ 2 - 4 ]. Few years later this vector
became very attractive for gene therapy, after the pseudotyping
with the G glycoprotein of Vesicular Stomatitis Virus (VSV-G)
[ 5 - 7 ]. VSV-G enveloped LVs can infect most cell types, are
particularly stable, and can be concentrated by ultracentrifugation
to titers exceeding 1 × 10 10 transducing unit (T.U.)/ml [ 8 ].
In contrast to other retroviral vectors, which can only transduce
actively replicating cells [ 9 ], LVs can transduce slowly or nondividing
cells [ 10 , 11 ], and hundreds of articles have been published on the
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