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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
11
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pubmed:dateCreated |
1995-12-1
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pubmed:abstractText |
We have developed novel self-inactivating and self-activating retroviral vectors based on the previously observed high-frequency deletion of direct repeats. We constructed spleen necrosis virus (SNV)-based viral vectors that contained large direct repeats flanking the viral encapsidation sequence (E). A large proportion of the proviruses in the target cells had E and one copy of the direct repeat deleted. Direct repeats of 1,333 and 788 bp were deleted at frequencies of 93 and 85%, respectively. To achieve a 100% deletion efficiency in target cells after ex vivo infection and drug selection, we constructed a self-activating vector that simultaneously deleted E and reconstituted the neomycin phosphotransferase gene. Selection of the target cells for resistance to G418 (a neomycin analog) ensured that all integrated proviruses had E deleted. The proviruses with E deleted were mobilized by a replication-competent virus 267,000-fold less efficiently than proviruses with E. We named these self-inactivating vectors E- (E-minus) vectors. These vectors should increase the safety of retroviral vector-mediated gene therapy by preventing the spread of vector sequences to nontarget cells in the event of coinfection with helper virus. We propose that direct-repeat deletions occur during RNA-dependent DNA synthesis and suggest that template switches occur without a requirement for RNA breaks. The minimum template dissociation frequency was estimated as 8%/100 bp per replication cycle. These vectors demonstrate that large direct repeats and template-switching properties of reverse transcriptase can be utilized to delete any sequence or reconstitute genes during retroviral replication.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-1329991,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-1372141,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-1608446,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-1696639,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-1702425,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-1729703,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-215703,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/7474097-8497071
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
69
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
6839-46
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:7474097-Animals,
pubmed-meshheading:7474097-Cell Line,
pubmed-meshheading:7474097-Cloning, Molecular,
pubmed-meshheading:7474097-Gene Therapy,
pubmed-meshheading:7474097-Genetic Vectors,
pubmed-meshheading:7474097-Humans,
pubmed-meshheading:7474097-Kanamycin Kinase,
pubmed-meshheading:7474097-Phosphotransferases (Alcohol Group Acceptor),
pubmed-meshheading:7474097-Proviruses,
pubmed-meshheading:7474097-RNA, Viral,
pubmed-meshheading:7474097-Recombinant Proteins,
pubmed-meshheading:7474097-Repetitive Sequences, Nucleic Acid,
pubmed-meshheading:7474097-Restriction Mapping,
pubmed-meshheading:7474097-Retroviridae,
pubmed-meshheading:7474097-Safety,
pubmed-meshheading:7474097-Sequence Deletion,
pubmed-meshheading:7474097-Transfection,
pubmed-meshheading:7474097-Virus Integration
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pubmed:year |
1995
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pubmed:articleTitle |
E- vectors: development of novel self-inactivating and self-activating retroviral vectors for safer gene therapy.
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