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Neuron-specific RNA interference using lentiviral vectors

Tolstrup Nielsen, Troels LU ; van Marion, Ingrid LU ; Hasholt, Lis and Lundberg, Cecilia LU (2009) In Journal of Gene Medicine 11(7). p.559-569
Abstract
Background Viral vectors have been used in several different settings for the delivery of small hairpin (sh) RNAs. However, most vectors have utilized ubiquitously-expressing polymerase (pol) III promoters to drive expression of the hairpin as a result Of the Strict requirement for precise transcriptional initiation and termination. Recently, pol 11 promoters have been used to construct vectors for RNA interference (RNAi). By embedding the shRNA into a micro RNA-context (miRNA) the endogenous miRNA processing machinery is exploited to achieve the mature synthetic miRNA (smiRNA), thereby expanding the possible promoter choices and eventually allowing cell type specific down-regulation of target genes. Methods In the present study, we... (More)
Background Viral vectors have been used in several different settings for the delivery of small hairpin (sh) RNAs. However, most vectors have utilized ubiquitously-expressing polymerase (pol) III promoters to drive expression of the hairpin as a result Of the Strict requirement for precise transcriptional initiation and termination. Recently, pol 11 promoters have been used to construct vectors for RNA interference (RNAi). By embedding the shRNA into a micro RNA-context (miRNA) the endogenous miRNA processing machinery is exploited to achieve the mature synthetic miRNA (smiRNA), thereby expanding the possible promoter choices and eventually allowing cell type specific down-regulation of target genes. Methods In the present study, we constructed lentiviral vectors expressing smiRNAs under the control of pol II promoters to knockdown gene expression in cell Culture and in the brain. Results We demonstrate robust kockdown of green fluorescent protein using lentiviral vectors driving RNAi from the ubiquitously-expressing promoter of the cytomegalovirus (CMV) and, in addition, we show for the first time neuron-specific knockdown in the brain using a neuron-specific promoter. Furthermore, we show that the expression pattern of the presumed ubiquitously-expressing CMV promoter changes over time from being expressed initially in neurons and glial cells to being expressed almost exclusively in neurons in later stages. Conclusions In the present study, we developed vectors for cell-specific RNAi for use in the brain. This offers the possibility of specifically targeting RNAi to a subset of cells in a complex tissue and may prove to be of great importance in the design of future gene therapeutic paradigms. Copyright (C) 2009 John Wiley & Sons, Ltd. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
lentiviral vectors, gene therapy, cell specificity, CNS, interference, RNA
in
Journal of Gene Medicine
volume
11
issue
7
pages
559 - 569
publisher
John Wiley & Sons
external identifiers
  • wos:000268113100002
  • scopus:68949213635
ISSN
1521-2254
DOI
10.1002/jgm.1333
language
English
LU publication?
yes
id
95140b05-92f3-4c4b-8d59-2e0d5d143515 (old id 1461520)
date added to LUP
2009-08-31 11:05:05
date last changed
2017-06-25 03:43:32
@article{95140b05-92f3-4c4b-8d59-2e0d5d143515,
  abstract     = {Background Viral vectors have been used in several different settings for the delivery of small hairpin (sh) RNAs. However, most vectors have utilized ubiquitously-expressing polymerase (pol) III promoters to drive expression of the hairpin as a result Of the Strict requirement for precise transcriptional initiation and termination. Recently, pol 11 promoters have been used to construct vectors for RNA interference (RNAi). By embedding the shRNA into a micro RNA-context (miRNA) the endogenous miRNA processing machinery is exploited to achieve the mature synthetic miRNA (smiRNA), thereby expanding the possible promoter choices and eventually allowing cell type specific down-regulation of target genes. Methods In the present study, we constructed lentiviral vectors expressing smiRNAs under the control of pol II promoters to knockdown gene expression in cell Culture and in the brain. Results We demonstrate robust kockdown of green fluorescent protein using lentiviral vectors driving RNAi from the ubiquitously-expressing promoter of the cytomegalovirus (CMV) and, in addition, we show for the first time neuron-specific knockdown in the brain using a neuron-specific promoter. Furthermore, we show that the expression pattern of the presumed ubiquitously-expressing CMV promoter changes over time from being expressed initially in neurons and glial cells to being expressed almost exclusively in neurons in later stages. Conclusions In the present study, we developed vectors for cell-specific RNAi for use in the brain. This offers the possibility of specifically targeting RNAi to a subset of cells in a complex tissue and may prove to be of great importance in the design of future gene therapeutic paradigms. Copyright (C) 2009 John Wiley & Sons, Ltd.},
  author       = {Tolstrup Nielsen, Troels and van Marion, Ingrid and Hasholt, Lis and Lundberg, Cecilia},
  issn         = {1521-2254},
  keyword      = {lentiviral vectors,gene therapy,cell specificity,CNS,interference,RNA},
  language     = {eng},
  number       = {7},
  pages        = {559--569},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Gene Medicine},
  title        = {Neuron-specific RNA interference using lentiviral vectors},
  url          = {http://dx.doi.org/10.1002/jgm.1333},
  volume       = {11},
  year         = {2009},
}