Journal article
Optimization of feline immunodeficiency virus vectors for RNA interference
Journal of virology, Vol.80(19), pp.9371-9380
10/01/2006
DOI: 10.1128/JVI.00958-06
PMCID: PMC1617215
PMID: 16973543
Abstract
RNA interference (RNAi) occurs naturally in plant and animal cells as a means for modulating gene expression. This process has been experimentally manipulated to achieve targeted gene silencing in cells, tissues, and animals, using a variety of vector systems. Here, we tested the hypothesis that vectors based on feline immunodeficiency virus (FIV) could be used for coexpression of reporter constructs and RNAi expression cassettes. We found, unexpectedly, in our initial constructs that placement of RNAi expression cassettes downstream from a polymerase II (pol II)-expressed reporter gene inhibited reporter expression but not vector titer. Through a series of intermediate vector constructs, we found that placement of the RNAi expression cassette relative to the Rev response element and the pol II expression cassette was critical for efficient RNAi and reporter gene expression. These results suggested that steric factors, including RNA structure and recruitment of competing transcriptional machinery, may affect gene expression from FIV vectors. In a second series of studies, we show that target sequence silencing can be achieved in cells transduced by FIV vectors coexpressing reporter genes and 3' untranslated region resident microRNAs. The optimized FIV-based RNAi expression vectors will find broad use given the extensive tropism of pseudotyped FIV vectors for many cell types in vitro and in vivo.
Details
- Title: Subtitle
- Optimization of feline immunodeficiency virus vectors for RNA interference
- Creators
- Scott Q Harper - University of IowaPatrick D Staber - University of IowaChristine R Beck - University of IowaSarah K Fineberg - University of IowaColleen Stein - University of IowaDalyz Ochoa - University of IowaBeverly L Davidson - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Journal of virology, Vol.80(19), pp.9371-9380
- DOI
- 10.1128/JVI.00958-06
- PMID
- 16973543
- PMCID
- PMC1617215
- NLM abbreviation
- J Virol
- ISSN
- 0022-538X
- eISSN
- 1098-5514
- Publisher
- American Society for Microbiology
- Grant note
- R01 HD044093 / NICHD NIH HHS F32 NS047048 / NINDS NIH HHS NS 047048 / NINDS NIH HHS T32 GM007337 / NIGMS NIH HHS HD 44093 / NICHD NIH HHS P30 DK054759 / NIDDK NIH HHS DK54759 / NIDDK NIH HHS
- Language
- English
- Date published
- 10/01/2006
- Academic Unit
- Internal Medicine; Ophthalmology and Visual Sciences
- Record Identifier
- 9984618515802771
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