Comparison of gene transfer efficiencies and gene expression levels achieved with equine infectious anemia virus- and human immunodeficiency virus type 1-derived lentivirus vectors.
Abstract: This report compares gene transfer efficiencies as well as durations and levels of gene expression for human immunodeficiency virus (HIV) and equine infectious anemia virus (EIAV) lentiviral vectors in a variety of human cell types in vitro. EIAV and HIV vectors transduced equivalent numbers of proliferating and G1/S- and G2/M-arrested cells, and both had very low efficiencies of transduction into G0-arrested cells. Analysis of the levels of both the enhanced green fluorescent protein (EGFP) and mRNA demonstrated that the HIV-transduced cells expressed greater levels of EGFP protein and RNA than the EIAV-transduced cells. Measurements of vector-derived EGFP RNA half-lives were fourfold higher with the HIV vector than with the EIAV vector. Long-term culture of EIAV-transduced human cells showed a significant decrease in the number of cells expressing the transgene; however, no corresponding loss was found in EIAV-transduced equine cells. In contrast, only a moderate decrease in the number of transgene-expressing cells was seen with the HIV vectors. Taken together, these results demonstrate that the EIAV vectors transduced human cells with efficiencies similar to those of the HIV vectors. However, our data indicate that transgene expression from EIAV vectors is limited by the instability of vector-derived RNA transcripts and silencing of the EIAV vectors over time.
Publication Date: 2002-01-05 PubMed ID: 11773424PubMed Central: PMC135783DOI: 10.1128/jvi.76.3.1510-1515.2002Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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This research explores the efficiency of gene transfer and expression when using the human immunodeficiency virus (HIV) and equine infectious anemia virus (EIAV) lentiviral vectors on various human cell types. The study revealed that while both vetor types showed comparable gene transfer efficiencies, the HIV vectors demonstrated higher levels of gene expression and long-term gene stability.
Research Method and Results
- The researchers tested the gene transfer efficiencies of HIV and EIAV vectors across various human cell types in a controlled laboratory setting.
- Both the HIV and EIAV vectors were able to transduce an equivalent number of proliferating cells and also G1/S- and G2/M-arrested cells (cells in different phase of cell cycle) implying that cell cycle stage does not impact the transduction efficiency of each vector.
- However, both vectors demonstrated low efficiency when transducing G0-arrested cells (non-dividing or quiescent stage cells).
- Next, the researchers looked at the levels of the enhanced green fluorescent protein (EGFP) and mRNA. They found that cells which had been transduced by HIV vectors expressed more EGFP protein and RNA than cells having been transduced with EIAV.
- The study revealed that the half-lives of vector-derived EGFP RNA were four times higher with the HIV vector compared to the EIAV vector.
Long-Term Culture and Transgene Expression
- The scientists observed that the long-term culture of EIAV-transduced human cells led to a significant decrease in the number of cells expressing the transgene, implying loss of gene expression over time.
- No such loss in gene expression was observed in EIAV-transduced equine cells, demonstrating a potential species-specific limitation.
- On the other hand, there was only a moderate decrease in the number of transgene-expressing cells long-term when the HIV vectors were used.
Conclusion
- Although both EIAV and HIV vectors demonstrated similar efficiencies in gene transfer, differences were observed in the level of transgene expression and its stability over time.
- The results suggest EIAV vectors have limited effectiveness due to the instability of vector-derived RNA transcripts and silencing of the vector over time.
- Therefore, HIV vectors might be a superior choice for gene therapy applications requiring sustained gene expression in human cells.
Cite This Article
APA
O'Rourke JP, Newbound GC, Kohn DB, Olsen JC, Bunnell BA.
(2002).
Comparison of gene transfer efficiencies and gene expression levels achieved with equine infectious anemia virus- and human immunodeficiency virus type 1-derived lentivirus vectors.
J Virol, 76(3), 1510-1515.
https://doi.org/10.1128/jvi.76.3.1510-1515.2002 Publication
Researcher Affiliations
- Children's Research Institute, Children's Hospital, Columbus, Ohio 43205, USA.
MeSH Terms
- Animals
- Cell Line
- Cell Line, Transformed
- Gene Expression
- Gene Transfer Techniques
- Genes, Reporter
- Genetic Vectors / genetics
- Green Fluorescent Proteins
- HIV-1 / genetics
- Horses
- Humans
- Infectious Anemia Virus, Equine / genetics
- Lentivirus / genetics
- Luminescent Proteins / genetics
- Luminescent Proteins / metabolism
- RNA Stability
- RNA, Messenger
- Time Factors
- Transduction, Genetic
- Tumor Cells, Cultured
Grant Funding
- F32 HL010430 / NHLBI NIH HHS
- R01 AI047693 / NIAID NIH HHS
- AI47693 / NIAID NIH HHS
- HL10430 / NHLBI NIH HHS
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