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Dynamics of transgene expression in a neural stem cell line transduced with lentiviral vectors incorporating the cHS4 insulator.

Jakobsson, Johan LU ; Rosenqvist, Nina LU ; Thompson, Lachlan LU ; Barraud, Perrine LU and Lundberg, Cecilia LU (2004) In Experimental Cell Research 298(2). p.611-623
Abstract
Transplantation of genetically manipulated cells to the central nervous system holds great promise for the treatment of several severe neurological disorders. The success of this strategy relies on sufficient levels of transgene expression after transplantation. This has been difficult to achieve, however, due to transgene silencing. In this study, we transduced the neural stem cell line RN33B with self-inactivating lentiviral vectors and analyzed transgenic expression of green fluorescent protein (GFP) in several different settings both in vitro and after transplantation to the brain. We found that the transgene was affected of silencing both when transduced cells were proliferating and after differentiation. To prevent silencing, the... (More)
Transplantation of genetically manipulated cells to the central nervous system holds great promise for the treatment of several severe neurological disorders. The success of this strategy relies on sufficient levels of transgene expression after transplantation. This has been difficult to achieve, however, due to transgene silencing. In this study, we transduced the neural stem cell line RN33B with self-inactivating lentiviral vectors and analyzed transgenic expression of green fluorescent protein (GFP) in several different settings both in vitro and after transplantation to the brain. We found that the transgene was affected of silencing both when transduced cells were proliferating and after differentiation. To prevent silencing, the cHS4 insulator was incorporated into the lentiviral vector. We found that a vector carrying the cHS4 insulator was partially protected against differentiation-dependent downregulation in vitro and in vivo. However, in proliferating cells, we found evidence for variegation and positional effects that were not prevented by the cHS4 insulator, suggesting that the mechanism behind silencing in proliferating cells is not the same mechanism influencing differentiation-dependent silencing. Taken together, these findings favor vector optimization as a strategy for achieving efficient ex vivo gene transfer in the central nervous system. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Gene therapy, Insulator, cHS4, Ex vivo, Silencing, Green fluorescent protein, Transplantation
in
Experimental Cell Research
volume
298
issue
2
pages
611 - 623
publisher
Academic Press
external identifiers
  • pmid:15265707
  • wos:000222985300026
  • scopus:3242682377
ISSN
1090-2422
DOI
10.1016/j.yexcr.2004.04.037
language
English
LU publication?
yes
id
da791337-d6e3-49be-b9d5-2806f2cd72ce (old id 125750)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15265707&dopt=Abstract
date added to LUP
2007-07-13 15:00:44
date last changed
2017-01-01 05:12:08
@article{da791337-d6e3-49be-b9d5-2806f2cd72ce,
  abstract     = {Transplantation of genetically manipulated cells to the central nervous system holds great promise for the treatment of several severe neurological disorders. The success of this strategy relies on sufficient levels of transgene expression after transplantation. This has been difficult to achieve, however, due to transgene silencing. In this study, we transduced the neural stem cell line RN33B with self-inactivating lentiviral vectors and analyzed transgenic expression of green fluorescent protein (GFP) in several different settings both in vitro and after transplantation to the brain. We found that the transgene was affected of silencing both when transduced cells were proliferating and after differentiation. To prevent silencing, the cHS4 insulator was incorporated into the lentiviral vector. We found that a vector carrying the cHS4 insulator was partially protected against differentiation-dependent downregulation in vitro and in vivo. However, in proliferating cells, we found evidence for variegation and positional effects that were not prevented by the cHS4 insulator, suggesting that the mechanism behind silencing in proliferating cells is not the same mechanism influencing differentiation-dependent silencing. Taken together, these findings favor vector optimization as a strategy for achieving efficient ex vivo gene transfer in the central nervous system.},
  author       = {Jakobsson, Johan and Rosenqvist, Nina and Thompson, Lachlan and Barraud, Perrine and Lundberg, Cecilia},
  issn         = {1090-2422},
  keyword      = {Gene therapy,Insulator,cHS4,Ex vivo,Silencing,Green fluorescent protein,Transplantation},
  language     = {eng},
  number       = {2},
  pages        = {611--623},
  publisher    = {Academic Press},
  series       = {Experimental Cell Research},
  title        = {Dynamics of transgene expression in a neural stem cell line transduced with lentiviral vectors incorporating the cHS4 insulator.},
  url          = {http://dx.doi.org/10.1016/j.yexcr.2004.04.037},
  volume       = {298},
  year         = {2004},
}