Document Detail


Transduction of Schistosoma japonicum schistosomules with vesicular stomatitis virus glycoprotein pseudotyped murine leukemia retrovirus and expression of reporter human telomerase reverse transcriptase in the transgenic schistosomes.
MedLine Citation:
PMID:  20692298     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Although draft genome sequences of two of the major human schistosomes, Schistosoma japonicum and Schistosoma mansoni are available, the structures and characteristics of most genes and the influence of exogenous genes on the metabolism of schistosomes remain uncharacterized. Furthermore, which functional genomics approaches will be tractable for schistosomes are not yet apparent. Here, the vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter, under the control of the retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce S. japonicum to investigate the utility of retrovirus-mediated transgenesis of S. japonicum and the activity of human telomerase reverse transcriptase as a reporter transgene in schistosomes. Schistosomules perfused from experimentally infected rabbits were cultured for 6 days after exposure to the virions after which genomic DNAs from virus exposed and control worms were extracted. Analysis of RNA from transduced parasites and immunohistochemistry of thin parasite sections revealed expression of hTERT in the transduced worms. Expression of hTERT was also confirmed by immunoblot analysis. These findings indicated that S. japonicum could be effectively transduced by VSVG-pseudotyped retrovirus carrying the hTERT gene. Given the potential of hTERT to aid in derivation of immortalized cells, these findings suggest that this pantropic retroviral approach can be employed to transduce cells from specific tissues and organs of schistosomes to investigate the influence of transgene hTERT on growth and proliferation of schistosome cells.
Authors:
Shenghui Yang; Paul J Brindley; Qingren Zeng; Yuesheng Li; Jun Zhou; Yan Liu; Biyuan Liu; Liting Cai; Tiebing Zeng; Qi Wei; Lingmei Lan; Donald P McManus
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-08-06
Journal Detail:
Title:  Molecular and biochemical parasitology     Volume:  174     ISSN:  1872-9428     ISO Abbreviation:  Mol. Biochem. Parasitol.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-10-05     Completed Date:  2011-02-03     Revised Date:  2013-12-19    
Medline Journal Info:
Nlm Unique ID:  8006324     Medline TA:  Mol Biochem Parasitol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  109-16     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier B.V. All rights reserved.
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MeSH Terms
Descriptor/Qualifier:
Animals
Animals, Genetically Modified / genetics,  metabolism*
Cell Line
Genes, Reporter
Humans
Immunoblotting
Immunohistochemistry
Leukemia Virus, Murine / genetics*,  metabolism
Membrane Glycoproteins / genetics*,  metabolism
Mice
NIH 3T3 Cells
Polymerase Chain Reaction
Rabbits
Reverse Transcriptase Polymerase Chain Reaction
Schistosoma japonicum / genetics,  growth & development,  metabolism,  virology*
Telomerase / genetics,  metabolism*
Transduction, Genetic*
Transgenes
Viral Envelope Proteins / genetics*,  metabolism
Grant Support
ID/Acronym/Agency:
R01 AI072773/AI/NIAID NIH HHS; R01AI072773/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/G protein, vesicular stomatitis virus; 0/Membrane Glycoproteins; 0/Viral Envelope Proteins; EC 2.7.7.49/TERT protein, human; EC 2.7.7.49/Telomerase
Comments/Corrections

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