Journal article
Inclusion of Jaagsiekte sheep retrovirus proviral elements markedly increases lentivirus vector pseudotyping efficiency
Molecular therapy, Vol.11(3), pp.460-469
03/2005
DOI: 10.1016/j.ymthe.2004.10.022
PMID: 15727943
Abstract
Retroviral pseudotyping for gene transfer applications endeavors to alter vector tropism and maintain a suitable titer. We investigated the compatibility of the Jaagsiekte sheep retrovirus (JSRV) envelope glycoprotein with the feline immunodeficiency virus (FIV) vector. A construct consisting of the minimal JSRV env coding region expressed from a standard mammalian expression plasmid generated FIV vector titers of approximately 10(4) TU/ml following standard triple transfection, collection of supernatants, and concentration by centrifuge. Interestingly, retention of the native proviral 5' and 3' flanking regions surrounding the JSRV env resulted in exceptional titers of approximately 10(8) TU/ml following the same viral preparation. To discern the regions necessary to achieve this 10,000-fold increase in titer, additional constructs were designed and tested. Our results indicate that the enhanced vector titer correlates with an increase in steady-state levels of envelope RNA that results from a combination of RNA splicing and stability, leading to increased envelope protein production. Expression of four other glycoproteins in an expression plasmid retaining the enhancing elements from the JSRV proviral sequence increased FIV vector titers from 0- to 100-fold. These novel data demonstrate that optimization of the envelope expression construct can profoundly influence titers for lentivirus vectors.
Details
- Title: Subtitle
- Inclusion of Jaagsiekte sheep retrovirus proviral elements markedly increases lentivirus vector pseudotyping efficiency
- Creators
- Patrick L Sinn - Department of Pediatrics, Program in Gene Therapy, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242, USAErin R BurnightHong ShenHung FanPaul B McCray Jr
- Resource Type
- Journal article
- Publication Details
- Molecular therapy, Vol.11(3), pp.460-469
- DOI
- 10.1016/j.ymthe.2004.10.022
- PMID
- 15727943
- NLM abbreviation
- Mol Ther
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Grant note
- HL-51670 / NHLBI NIH HHS R01 HL-61460 / NHLBI NIH HHS P30 DK-54759 / NIDDK NIH HHS HL-67623 / NHLBI NIH HHS
- Language
- English
- Date published
- 03/2005
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine; Ophthalmology and Visual Sciences
- Record Identifier
- 9984093370702771
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