1998 Fiscal Year Final Research Report Summary
Development of a novel retrovirus vector system for hematopoietic Stem cells
| Project/Area Number |
09671106
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
ITOH Katsuhiko Kyoto University, Graduate school of medicine, Lectu, 医学研究科, 講師 (90281097)
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| Co-Investigator(Kenkyū-buntansha) |
KANEKO Yoshiyuki Kyoto University, Graduate School of Medicine Assistant Professor, 医学研究科, 助手 (40252465)
FUJITA Jun Kyoto University, Graduate School of Medicine, Profess, 医学研究科, 教授 (50173430)
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| Project Period (FY) |
1997 – 1998
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| Keywords | gene therapy / hematopoietic stem cell / retrovirus |
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| Research Abstract |
The aims of the project was as follows ;
1. Evaluation of the gene expression by a novel retroviral vector which consisted of the SFFVp LTR and the MESV leader sequences.
2. Establishment and the evaluation of novel packaging cells based on stromal cells.
3. Evaluation of the pseudo-typed retroviral vectors based on SFFVp with VSV G-protein.
We have obtained the following results ;
1. A series of retroviral vectors were constructed. Using the packaging cells based on NIH/3T3, these vector-producing cells were isolated. Expression levels of these vectors were compared in hematopoietic progenitors. The highest expression of a marker gene among these was obtained by the FMEV-type vector, which contained the SFFVp LTR and the MESV leader sequences.
2. Plasmid DNAs of the expression vector carrying gal/pol sequences and that carrying the MoMLV env sequences were introduced into a stromal cell line, MS-5. These novel stromal cell-based packaging cells were found to be unstable to produce high titer of viral vectors.
3. SFFVp based retroviral vectors could be pseudo-typed with VSV G-protein and these particles successfully transduced the cells.
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