Journal article
rAAV2 Traffics through both the Late and the Recycling Endosomes in a Dose-Dependent Fashion
Molecular therapy, Vol.13(4), pp.671-682
04/2006
DOI: 10.1016/j.ymthe.2005.12.002
PMCID: PMC1712670
PMID: 16442847
Abstract
Inefficient trafficking of recombinant adeno-associated virus type-2 (rAAV2) to the nucleus is a major barrier for transduction. Using imaging and subcellular fractionation techniques, we evaluated the extent of rAAV2 movement through the late (Rab7) and recycling (Rab11) endosomes. Following rAAV2 infection of HeLa cells, immunoisolation of HA-Rab7- or HA-Rab11-tagged endosomes and intracellular colocalization of Cy3-labeled rAAV2 with EGFP-Rab7 or EGFP-Rab11 markers demonstrated dose-dependent trafficking of rAAV2 through the recycling and late endosomal compartments. At low multiplicities of infection (m.o.i. 100 genomes/cell), rAAV2 predominantly trafficked to the Rab7 compartment. In contrast, rAAV2 predominantly trafficked to the recycling endosome at 100-fold higher m.o.i. siRNA studies inhibiting either Rab7 or Rab11 demonstrated that reducing Rab11 protein levels more significantly inhibited rAAV2 transduction on a per genome basis compared to inhibition of Rab7. Dose-response curves, comparing the m.o.i. of AV2Luc infection to relative transduction, also supported the hypothesis that viral movement through the Rab11 compartment at high m.o.i. is more competent for transgene expression (∼100-fold) than virus that moves through the Rab7 compartment at low m.o.i. These findings suggest that strategies to shunt viral movement from the late to the recycling endosome may be effective at increasing viral transduction for gene therapy.
Details
- Title: Subtitle
- rAAV2 Traffics through both the Late and the Recycling Endosomes in a Dose-Dependent Fashion
- Creators
- Wei Ding - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USALiang N Zhang - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USACharles Yeaman - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USAJohn F Engelhardt - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USA
- Resource Type
- Journal article
- Publication Details
- Molecular therapy, Vol.13(4), pp.671-682
- DOI
- 10.1016/j.ymthe.2005.12.002
- PMID
- 16442847
- PMCID
- PMC1712670
- NLM abbreviation
- Mol Ther
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Language
- English
- Date published
- 04/2006
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984025462102771
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