Abstract
Gene therapy in eukaryotes has met many obstacles. Research into the design of suitable nonviral vectors has been slow. To our knowledge, no nonviral vector has been proposed that allows for the possibility of highly efficient, site-selective integration into the genome of mammalian cells. On the basis of prior studies investigating the components necessary for transposon, retrovirus-like retrotransposon, and retroviral integration, we propose a nonviral system that would potentially allow for site-selective, efficient integration into the mammalian genome. Transposons have been developed that can transform a variety of cell lines. For example, the Sleeping Beauty transposon (SB) can transform a wide range of vertebrate cells from fish to human, and it mediates stable integration and long-term transgene expression in mice. However, the efficiency of transposition varies significantly among cell lines, suggesting the possible involvement of host factors in SB transposition. Here, we propose the use of a chimeric transposase (i.e., transposase-host DNA binding domain) to bypass the potential requirement of a host DNA-directing factor (or factors) for efficient, site-selective integration. We also discuss another potential method of docking the transposon-based vector adjacent to the host DNA, utilizing repetitive sequences for homologous recombination to promote efficient site-selective integration, as well as other site-selective nonviral approaches.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1242-1247 |
| Number of pages | 6 |
| Journal | FASEB Journal |
| Volume | 16 |
| Issue number | 10 |
| DOIs | |
| State | Published - Aug 7 2002 |
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Keywords
- Gene therapy
- Recombination
- Transposon
ASJC Scopus subject areas
- Biotechnology
- Biochemistry
- Molecular Biology
- Genetics
Cite this
Design of a nonviral vector for site-selective, efficient integration into the human genome. / Kaminski, Joseph Michael; Huber, Mark R.; Summers, James B.; Ward, Matthew B.
In: FASEB Journal, Vol. 16, No. 10, 07.08.2002, p. 1242-1247.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Design of a nonviral vector for site-selective, efficient integration into the human genome
AU - Kaminski, Joseph Michael
AU - Huber, Mark R.
AU - Summers, James B.
AU - Ward, Matthew B.
PY - 2002/8/7
Y1 - 2002/8/7
N2 - Gene therapy in eukaryotes has met many obstacles. Research into the design of suitable nonviral vectors has been slow. To our knowledge, no nonviral vector has been proposed that allows for the possibility of highly efficient, site-selective integration into the genome of mammalian cells. On the basis of prior studies investigating the components necessary for transposon, retrovirus-like retrotransposon, and retroviral integration, we propose a nonviral system that would potentially allow for site-selective, efficient integration into the mammalian genome. Transposons have been developed that can transform a variety of cell lines. For example, the Sleeping Beauty transposon (SB) can transform a wide range of vertebrate cells from fish to human, and it mediates stable integration and long-term transgene expression in mice. However, the efficiency of transposition varies significantly among cell lines, suggesting the possible involvement of host factors in SB transposition. Here, we propose the use of a chimeric transposase (i.e., transposase-host DNA binding domain) to bypass the potential requirement of a host DNA-directing factor (or factors) for efficient, site-selective integration. We also discuss another potential method of docking the transposon-based vector adjacent to the host DNA, utilizing repetitive sequences for homologous recombination to promote efficient site-selective integration, as well as other site-selective nonviral approaches.
AB - Gene therapy in eukaryotes has met many obstacles. Research into the design of suitable nonviral vectors has been slow. To our knowledge, no nonviral vector has been proposed that allows for the possibility of highly efficient, site-selective integration into the genome of mammalian cells. On the basis of prior studies investigating the components necessary for transposon, retrovirus-like retrotransposon, and retroviral integration, we propose a nonviral system that would potentially allow for site-selective, efficient integration into the mammalian genome. Transposons have been developed that can transform a variety of cell lines. For example, the Sleeping Beauty transposon (SB) can transform a wide range of vertebrate cells from fish to human, and it mediates stable integration and long-term transgene expression in mice. However, the efficiency of transposition varies significantly among cell lines, suggesting the possible involvement of host factors in SB transposition. Here, we propose the use of a chimeric transposase (i.e., transposase-host DNA binding domain) to bypass the potential requirement of a host DNA-directing factor (or factors) for efficient, site-selective integration. We also discuss another potential method of docking the transposon-based vector adjacent to the host DNA, utilizing repetitive sequences for homologous recombination to promote efficient site-selective integration, as well as other site-selective nonviral approaches.
KW - Gene therapy
KW - Recombination
KW - Transposon
UR - http://www.scopus.com/inward/record.url?scp=0036323154&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036323154&partnerID=8YFLogxK
U2 - 10.1096/fj.02-0127hyp
DO - 10.1096/fj.02-0127hyp
M3 - Article
VL - 16
SP - 1242
EP - 1247
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 10
ER -